Brendan+C.

__**Week of 12/1/13-12/7/13**__

Figure: FPLC spec results. Tubes collected = 39-48 for reconcentration.

Figure: PreFPLC spec results after Ni-NTA assay of new batch of protein NDM-1.

Figure 1: Verification assay of protein activity after new batch of NDM-1 made.

Figure: Image of characterization of protein solutions at different stages of purification.

Analysis: We now have enough protein to do 4,500 spec analyses for assays. We've shown that the protein solution is more pure and that the activity is present. FPLC results were quite good and initial protein concentration post Ni-NTA column was very high. The gel indicates very good purity of protein in the FPLC lane. Note that the sample from FPLC was diluted prior to adding to the gel due to the high concentration of protein.

__**Week of 11/24-11/30/13**__

Figure: PreFPLC spec results after Ni-NTA assay of new batch of protein NDM-1.

Very high concentration of protein. Not sure if all of it is the target protein however. Gel characterization indicates a lot of contaminating protein so FPLC will be used to get rid of it.

__**Week of 11/17/13-11/23/13**__

//11/23/13//: Final attempt at activity assay. - @ 490nm __Activity Assay:__ 0.25ug in 10uL buffer (so add 19uL buffer per 1uL solution) Dilute to 20uM nitrocefin in 0.5mL soln in cuvette 50mM Hepes, 10uM ZnSO4. 

__Inhibition Assay:__

//11/19/13// - HF9 plates GOLD results are in.


 * Table 1:** GOLD docking results of 4EY2 vs HF9 plates (~40,000 ligands). Top 30 scores shown.
 * # || Score || S(PLP) || S(hbond) || S(cho) || S(metal) || DE(clash) || DE(tors) || intcor || time || name ||
 * 1 || 102.49 || -77.9 || 1.88 || 0 || 3.96 || 0 || 2.62 || 0.41 || 44.488 || 'KM10420SC' ||
 * 2 || 100.93 || -89.68 || 0.6 || 0 || 2 || 0 || 2.55 || 2.55 || 139.309 || 'SPB06607SC' ||
 * 3 || 100.64 || -65.71 || 2.79 || 0 || 4.76 || 0 || 1.86 || 1.75 || 141.639 || 'CD09635SC' ||
 * 4 || 99.31 || -91.4 || 0.65 || 1 || 1 || 0.31 || 3.3 || 3.26 || 192.226 || 'HTS11951SC' ||
 * 5 || 98.36 || -75.17 || 4.69 || 0 || 2.1 || 0 || 2.23 || 0.99 || 66.862 || 'NRB05197SC' ||
 * 6 || 97.81 || -74.75 || 2.65 || 0 || 2.87 || 0 || 1.86 || 1.63 || 196.913 || 'BTB13323SC' ||
 * 7 || 97.65 || -84.3 || 0 || 1 || 1.88 || 0 || 1.79 || 2.65 || 21.541 || 'HTS08270SC' ||
 * 8 || 97.54 || -81.81 || 1 || 0 || 2.63 || 0 || 2.16 || 0.99 || 28.486 || 'HTS09917SC' ||
 * 9 || 97.32 || -81.04 || 1 || 0 || 2.94 || 0 || 3.46 || 2.58 || 176.638 || 'HTS02114SC' ||
 * 10 || 96.63 || -79.59 || 0.88 || 0 || 2.97 || 0.18 || 3.18 || 3.01 || 92.801 || 'RH01689SC' ||
 * 11 || 96.52 || -76.77 || 0 || 0 || 3.94 || 0 || 2.08 || 0.13 || 28.004 || 'SCR00160SC' ||
 * 12 || 96.27 || -99.1 || 0.43 || 0 || 0 || 0 || 2.16 || 0.2 || 87.649 || 'HTS07745SC' ||
 * 13 || 95.73 || -80.39 || 0.8 || 0 || 2.86 || 0 || 3.22 || 2.19 || 116.015 || 'HTS08667SC' ||
 * 14 || 95.71 || -80.05 || 2 || 0 || 1.99 || 0 || 2.26 || 2.22 || 142.861 || 'NRB01104SC' ||
 * 15 || 95.53 || -80.37 || 0.9 || 0 || 2.92 || 0.58 || 2.3 || 0.13 || 85.736 || 'HTS08664SC' ||
 * 16 || 95.35 || -67.09 || 2.93 || 0 || 3.89 || 0 || 2.56 || 1.14 || 116.92 || 'HTS08000SC' ||
 * 17 || 94.86 || -72.23 || 1.99 || 0 || 3 || 0 || 0.73 || 0.12 || 115.001 || 'HTS08318SC' ||
 * 18 || 94.85 || -73.47 || 2.71 || 0 || 2.94 || 0 || 4.44 || 4.53 || 141.635 || 'HTS02720SC' ||
 * 19 || 94.84 || -68.78 || 1.38 || 1 || 3.5 || 0.48 || 1 || 0.38 || 36.759 || 'HTS01400SC' ||
 * 20 || 94.84 || -86.28 || 0 || 0 || 2 || 0 || 1.98 || 0.52 || 60.642 || 'CD08739SC' ||
 * 21 || 94.78 || -80.72 || 0 || 0 || 2.83 || 0 || 1.54 || 0.13 || 111.982 || 'CD12049SC' ||
 * 22 || 94.76 || -78 || 0.8 || 0 || 2.66 || 0 || 1.78 || 1.95 || 18.199 || 'HTS12773SC' ||
 * 23 || 94.18 || -73.09 || 0 || 0 || 3.99 || 0 || 1.55 || 0.18 || 24.803 || 'HTS01661SC' ||
 * 24 || 94.17 || -77.29 || 0 || 0 || 3.44 || 0.15 || 3 || 2.41 || 21.914 || 'NRB04246SC' ||
 * 25 || 93.95 || -81.61 || 0.99 || 0 || 2 || 0 || 1.85 || 1.09 || 64.839 || 'SPB03389SC' ||
 * 26 || 93.84 || -77.69 || 0 || 0 || 2.94 || 0 || 1.86 || 2.25 || 30.367 || 'SEW05237SC' ||
 * 27 || 93.68 || -73.59 || 3.63 || 0 || 1.97 || 0 || 2.46 || 2.3 || 129.884 || 'HTS11140SC' ||
 * 28 || 93.65 || -79.83 || 0.5 || 0 || 2.65 || 0 || 2.98 || 2.25 || 113.86 || 'SPB01505SC' ||
 * 29 || 93.38 || -76.64 || 0 || 0 || 3.67 || 0 || 2.76 || 0.2 || 22.886 || 'HTS00696SC' ||
 * 30 || 93.35 || -71.81 || 1.32 || 1 || 2.96 || 0 || 2.71 || 2.26 || 129.794 || 'HTS05874SC' ||

__**Week of 11/10/13-11/16/13**__

//11/15/13//: Protein Purification and Characterization
 * Figure 1:** FPLC results of purification of second batch of NDM-1.

In short, the FPLC seems to have worked. We took sample tubes 41-46 and reconcentrated to 1mL. The resulting solution was nanodropped and determined to be 0.58mg/mL. This was conducted on a batch of protein on which expression was begun earlier in the week. After FPLC and concentration was completed, snapfreezing was conducted.

Meanwhile, another batch of protein expression is occurring. To this batch, Ni-NTA column purification was conducted.

These results show significantly less protein in the elution 2 after changing wash imidazole concentration to 15mM.

//11/12/13//: Protein purification
 * Figure 1:** Nanodrop of elution 1 after Ni-NTA purification.

Analysis: No distinct 280 peak suggests a not-so-great protein expression. The high 230 peak suggests considerable contamination. Some things to consider for the future = FPLC and rerunning the Ni-NTA column. 0.56mg/mL is not a terrible concentration however, though repurification will most likely be coming up.



Indicates reason to use FPLC to purify batch 1.
 * Figure 3:** Batch 1 of protein made after Ni-NTA purification. Lane 1 indicates Elution 1 and lane 2 indicates Elution 2.

The gel for batch 2 is currently destaining after Ni-NTA purification.

//11/11/13//: Protein expression continuing.


 * Table 1:** OD600 to 0.5 abs data prior to IPTG induction (set 2)
 * Time || mL added of small culture || Large culture 1 OD600 || Large culture 2 OD600 ||
 * 2:14 || 0 || 0 ||  ||
 * || 5 || 0 ||  ||
 * || 10 || 0.021 ||  ||
 * || 15 || 0.08 ||  ||
 * || 18 || 0.1 || 0.16 ||
 * 3:18 ||  || 0.12 || 0.2 ||
 * 3:48 ||  || 0.29 || 0.478 ||
 * 3:58 ||  || 0.438 || 0.544 ||
 * 4:13 ||  || 0.567 ||   ||

__**Week of 11/3/13-11/9/13**__

//11/8/13//: 4EY2 vs HF9plates screening still incomplete on the DDFE. Protein expression continuing.


 * Table 1:** OD600 to 0.5 absorbance data prior to IPTG induction.


 * Flask || Time || OD(600) || Flask 2 || Time || OD600 ||
 * 1 ||  2:09  ||  0.099  ||  2  ||  2:14  ||  0.13  ||
 * Added to incubator ||  ||   ||   ||   ||   ||
 * || 3:05  ||  0.087  ||   || 3:07PM ||  0.08  ||
 * || 330  ||  0.101  ||   ||  330  ||  0.1  ||
 * || 412  ||  0.181  ||   ||  412  ||  0.209  ||
 * || 4:52  ||  0.369  ||   ||  4:46  ||  0.422  ||
 * || 5:04  ||  0.452  ||   ||  4:56  ||  0.45  ||
 * || 5:10  ||  0.503  ||   ||  5:05  ||  0.51  ||
 * || 5:10  ||  0.503  ||   ||  5:05  ||  0.51  ||

//11/6/13:// 4EY2 vs HF9plates screening begun. -Protein expression begun - small scale culture

//11/5/13:// Screened 4EY2 vs CBDiversity in GOLD. Plates of BL21(DE3) transformed with pet27b+NDM1-betalactamase made.


 * Table 1:** 4EY2 docked to CBDiversity in GOLD (top 30 shown).


 * # || Score || S(PLP) || S(hbond) || S(cho) || S(metal) || DE(clash) || DE(tors) || intcor || time || name ||
 * 1 ||  109.78  ||  -50.03  ||  0.4  ||  0  ||  9.89  ||  0  ||  0.57  ||  0.32  ||  18.27  || '5587264' ||
 * 2 ||  109.44  ||  -62.12  ||  3.27  ||  0  ||  6.73  ||  0  ||  1.56  ||  0.25  ||  85.953  || '7784870' ||
 * 3 ||  108.17  ||  -81.24  ||  0.02  ||  0  ||  4.9  ||  0  ||  1.82  ||  1.12  ||  95.191  || '7638084' ||
 * 4 ||  107.91  ||  -85.25  ||  1  ||  0  ||  3.77  ||  0  ||  1.79  ||  0.44  ||  112.468  || '5321217' ||
 * 5 ||  107.4  ||  -40.78  ||  3  ||  0  ||  9.9  ||  0  ||  2.26  ||  2.32  ||  16.458  || '5238378' ||
 * 6 ||  107.2  ||  -67.07  ||  1.92  ||  0  ||  5.85  ||  0  ||  0.78  ||  0.62  ||  97.993  || '7565278' ||
 * 7 ||  107.18  ||  -70.79  ||  0.78  ||  0  ||  5.96  ||  0.01  ||  1.29  ||  0.72  ||  93.248  || '7587987' ||
 * 8 ||  106.84  ||  -70.95  ||  1.92  ||  0  ||  5.7  ||  0  ||  3.29  ||  2.45  ||  94.92  || '7746383' ||
 * 9 ||  106.48  ||  -65.93  ||  2.87  ||  0  ||  5.95  ||  0  ||  2.36  ||  0.92  ||  76.339  || '6639676' ||
 * 10 ||  106.36  ||  -66.66  ||  2.42  ||  0  ||  5.93  ||  0  ||  1.89  ||  0.48  ||  37.107  || '5378944' ||
 * 11 ||  106.24  ||  -87.06  ||  0  ||  0  ||  3.88  ||  0.41  ||  3.97  ||  3.57  ||  133.13  || '6462982' ||
 * 12 ||  106.12  ||  -56.24  ||  3  ||  0  ||  7.67  ||  0  ||  2.75  ||  0.34  ||  65.994  || '5157370' ||
 * 13 ||  106.07  ||  -81.31  ||  1  ||  0  ||  3.98  ||  0  ||  1.14  ||  0.17  ||  98.259  || '7009439' ||
 * 14 ||  105.8  ||  -67.86  ||  2.15  ||  0  ||  5.88  ||  0.08  ||  1.99  ||  0.14  ||  118.286  || '5378829' ||
 * 15 ||  105.53  ||  -74.01  ||  0  ||  1  ||  5.73  ||  0.38  ||  3.29  ||  0.92  ||  144.458  || '6457101' ||
 * 16 ||  105.5  ||  -65.38  ||  1.94  ||  0  ||  5.85  ||  0  ||  0.74  ||  0.57  ||  55.16  || '7584104' ||
 * 17 ||  105.46  ||  -61.13  ||  0  ||  0  ||  7.87  ||  0  ||  2.54  ||  2.18  ||  24.301  || '9048987' ||
 * 18 ||  105.36  ||  -64.75  ||  2.54  ||  0  ||  5.91  ||  0  ||  1.52  ||  0.58  ||  33.022  || '5378880' ||
 * 19 ||  105.31  ||  -59.56  ||  0  ||  0  ||  7.99  ||  2.64  ||  1.63  ||  3.73  ||  29.417  || '5538434' ||
 * 20 ||  105.28  ||  -74.85  ||  0.17  ||  0  ||  5.59  ||  2.41  ||  2.77  ||  4.33  ||  138.844  || '7722178' ||
 * 21 ||  105.16  ||  -65.55  ||  1.67  ||  0  ||  5.97  ||  0  ||  0.67  ||  0.11  ||  29.696  || '7644832' ||
 * 22 ||  105.12  ||  -60.2  ||  0  ||  0  ||  7.73  ||  0  ||  0.76  ||  0.06  ||  52.8  || '6052424' ||
 * 23 ||  105.1  ||  -59.88  ||  0  ||  0  ||  7.85  ||  0  ||  0.98  ||  0.06  ||  34.304  || '6052410' ||
 * 24 ||  104.94  ||  -64.67  ||  2  ||  0  ||  5.88  ||  10.48  ||  0.96  ||  11.38  ||  147.529  || '7551148' ||
 * 25 ||  104.76  ||  -59.9  ||  0  ||  0  ||  7.82  ||  0  ||  1.05  ||  0.06  ||  51.932  || '6052398' ||
 * 26 ||  104.7  ||  -76.95  ||  2.08  ||  0  ||  3.96  ||  1.21  ||  1.82  ||  2.4  ||  98.638  || '7589134' ||
 * 27 ||  104.36  ||  -77  ||  0  ||  0  ||  5.61  ||  0  ||  3.27  ||  0.23  ||  80.345  || '7583099' ||
 * 28 ||  104.33  ||  -81.93  ||  0  ||  1  ||  3.88  ||  0  ||  3.32  ||  2.77  ||  39.927  || '7632139' ||
 * 29 ||  104.28  ||  -72.6  ||  0.97  ||  0  ||  4.88  ||  0  ||  1.06  ||  1.6  ||  77.234  || '6422125' ||
 * 30 ||  104.22  ||  -58.16  ||  0  ||  0  ||  7.91  ||  0.9  ||  2.26  ||  3.68  ||  22.215  || '5226803' ||

__**Week of 10/27/13-11/2/13**__

// 10/31/13: // 4EY2 Docking to CB306 in GOLD


 * Table 1:** 4EY2 docked to CB306 in GOLD.
 * # || Score || S(PLP) || S(hbond) || S(cho) || S(metal) || DE(clash) || DE(tors) || intcor || time || name ||
 * 1 ||  44.12  ||  -39.11  ||  2  ||  0  ||  0  ||  0  ||  1.7  ||  2.4  ||  4.123  || '5739880' ||
 * 2 ||  41.77  ||  -22.9  ||  6.62  ||  0  ||  0  ||  0  ||  0.96  ||  0.33  ||  4.425  || '7726180' ||
 * 3 ||  40.48  ||  -36.14  ||  1.12  ||  0.81  ||  0  ||  0  ||  0.8  ||  0.14  ||  4.472  || '9027311' ||
 * 4 ||  40.33  ||  -28  ||  5  ||  0  ||  0  ||  0  ||  1.42  ||  0.16  ||  4.122  || '7575772' ||
 * 5 ||  40.21  ||  -35.49  ||  1.67  ||  0  ||  0  ||  0  ||  0.4  ||  0.41  ||  4.142  || '7951649' ||
 * 6 ||  39.89  ||  -32.68  ||  2.91  ||  0  ||  0  ||  0  ||  0.81  ||  0.09  ||  3.261  || '5978818' ||
 * 7 ||  39.53  ||  -39.77  ||  0  ||  0  ||  0  ||  0  ||  0.61  ||  0.97  ||  4.059  || '5627579' ||
 * 8 ||  39.38  ||  -31.46  ||  3  ||  0  ||  0  ||  0  ||  0.55  ||  0.01  ||  1.761  || '9040855' ||
 * 9 ||  38.86  ||  -34.19  ||  2.63  ||  0  ||  0  ||  0  ||  1.79  ||  0.11  ||  3.507  || '7543758' ||
 * 10 ||  38.83  ||  -31.2  ||  2.78  ||  0  ||  0  ||  0  ||  0.37  ||  0.01  ||  2.827  || '5761107' ||
 * 11 ||  38.73  ||  -34.5  ||  2  ||  0  ||  0  ||  0.01  ||  0.98  ||  0.15  ||  5.063  || '7746693' ||
 * 12 ||  38.71  ||  -33.31  ||  2.34  ||  0  ||  0  ||  0  ||  2.33  ||  3.04  ||  4.02  || '7623092' ||
 * 13 ||  38.63  ||  -32.39  ||  3.13  ||  0  ||  0  ||  0  ||  1.63  ||  0.07  ||  3.724  || '7814849' ||
 * 14 ||  38.53  ||  -37.81  ||  1.36  ||  0  ||  0  ||  0  ||  1.79  ||  0.23  ||  4.078  || '7938694' ||
 * 15 ||  38.13  ||  -27.15  ||  4  ||  0.83  ||  0  ||  0  ||  3.05  ||  2.61  ||  0.958  || '6629394' ||
 * 16 ||  38.12  ||  -34.02  ||  1.66  ||  0  ||  0  ||  0  ||  0.89  ||  0.54  ||  5.123  || '5654317' ||
 * 17 ||  38.11  ||  -33.11  ||  1.8  ||  0  ||  0  ||  0  ||  0.21  ||  0.02  ||  2.869  || '6664007' ||
 * 18 ||  38.04  ||  -31.28  ||  2.78  ||  0  ||  0  ||  0  ||  0.81  ||  0.01  ||  3.924  || '5957448' ||
 * 19 ||  37.93  ||  -30.82  ||  2.93  ||  0  ||  0  ||  0  ||  0.84  ||  0  ||  0.964  || '9061037' ||
 * 20 ||  37.83  ||  -31.33  ||  2.6  ||  0  ||  0  ||  0  ||  1.34  ||  0.86  ||  4.983  || '7558664' ||
 * 21 ||  37.7  ||  -32.81  ||  1.98  ||  0  ||  0  ||  0  ||  0.91  ||  0.78  ||  4.577  || '5525637' ||
 * 22 ||  37.7  ||  -36.33  ||  0.74  ||  0  ||  0  ||  0  ||  0.43  ||  0.02  ||  4.64  || '5728150' ||
 * 23 ||  37.62  ||  -21.09  ||  6  ||  0  ||  0  ||  0  ||  0.91  ||  0.24  ||  3.036  || '5467784' ||
 * 24 ||  37.57  ||  -21.33  ||  6.24  ||  0  ||  0  ||  0  ||  1.44  ||  0.1  ||  3.472  || '5192853' ||
 * 25 ||  37.46  ||  -33.34  ||  1.81  ||  0  ||  0  ||  0  ||  0.95  ||  0.52  ||  3.41  || '5241283' ||
 * 26 ||  37.26  ||  -36.94  ||  0.98  ||  0  ||  0  ||  0  ||  1.37  ||  0.12  ||  2.984  || '7766330' ||
 * 27 ||  37.25  ||  -32.89  ||  2  ||  0  ||  0  ||  0  ||  0.86  ||  0.08  ||  3.654  || '7724431' ||
 * 28 ||  37.19  ||  -32.08  ||  2.18  ||  0  ||  0  ||  0  ||  0.85  ||  0.28  ||  3.42  || '7698680' ||
 * 29 ||  36.98  ||  -27.47  ||  3.76  ||  0  ||  0  ||  0  ||  1.95  ||  2.09  ||  3.615  || '7676009' ||
 * 30 ||  36.96  ||  -36.31  ||  1  ||  0  ||  0  ||  0  ||  1.32  ||  0.28  ||  4.869  || '7930734' ||
 * 31 ||  36.86  ||  -33.22  ||  1.99  ||  0  ||  0  ||  0  ||  1.19  ||  0.03  ||  2.73  || '5531413' ||
 * 32 ||  36.79  ||  -29.28  ||  2.99  ||  0  ||  0  ||  0  ||  0.74  ||  0.02  ||  3.6  || '7935514' ||
 * 33 ||  36.58  ||  -23.03  ||  4.92  ||  0  ||  0  ||  0  ||  0.69  ||  0.1  ||  3.613  || '5282931' ||
 * 34 ||  36.56  ||  -30.22  ||  2.98  ||  0.35  ||  0  ||  0  ||  1.83  ||  0.01  ||  2.29  || '9045960' ||
 * 35 ||  36.38  ||  -32.31  ||  1.57  ||  0  ||  0  ||  0  ||  0.33  ||  0.02  ||  3.36  || '5158497' ||
 * 36 ||  35.76  ||  -36.72  ||  0.06  ||  0  ||  0  ||  0  ||  1.23  ||  1.32  ||  2.673  || '9038774' ||
 * 37 ||  35.75  ||  -28.78  ||  2.74  ||  0  ||  0  ||  0  ||  0.63  ||  0.01  ||  1.985  || '7575112' ||
 * 38 ||  35.65  ||  -28.48  ||  2.61  ||  0  ||  0  ||  0  ||  0.35  ||  0.05  ||  3.216  || '7741517' ||
 * 39 ||  35.64  ||  -34.55  ||  1.58  ||  0  ||  0  ||  0  ||  1.84  ||  0.01  ||  2.2  || '7948268' ||
 * 40 ||  35.62  ||  -32.98  ||  1.38  ||  0  ||  0  ||  0  ||  0.77  ||  0.03  ||  3.843  || '5193892' ||
 * 41 ||  35.58  ||  -31.43  ||  1.55  ||  0  ||  0  ||  0  ||  0.25  ||  0.03  ||  2.64  || '7437091' ||
 * 42 ||  35.52  ||  -31.42  ||  1.96  ||  0  ||  0  ||  0  ||  0.92  ||  0.04  ||  2.414  || '7608656' ||
 * 43 ||  35.39  ||  -30.63  ||  1.96  ||  0  ||  0  ||  0  ||  0.62  ||  0.07  ||  4.068  || '9062573' ||
 * 44 ||  35.31  ||  -34.26  ||  0.99  ||  0  ||  0  ||  0  ||  1.71  ||  1.49  ||  3.268  || '7911350' ||
 * 45 ||  35.17  ||  -31.54  ||  2  ||  0  ||  0  ||  0.01  ||  1.94  ||  1.52  ||  2.783  || '5301198' ||
 * 46 ||  35.1  ||  -19.45  ||  5.68  ||  0  ||  0  ||  0  ||  0.69  ||  0.02  ||  2.688  || '6433056' ||
 * 47 ||  35.09  ||  -34.17  ||  0.51  ||  0  ||  0  ||  0  ||  0.32  ||  0.03  ||  2.53  || '7870889' ||
 * 48 ||  35.06  ||  -32.57  ||  1.54  ||  0  ||  0  ||  0  ||  1.42  ||  0.72  ||  6.202  || '6956655' ||
 * 49 ||  34.97  ||  -29.09  ||  2  ||  0  ||  0  ||  0  ||  0.07  ||  0.02  ||  2.739  || '7829885' ||
 * 50 ||  34.96  ||  -30.63  ||  1.59  ||  0  ||  0  ||  0  ||  0.21  ||  0  ||  2.61  || '5215541' ||
 * 51 ||  34.79  ||  -28.9  ||  2  ||  0  ||  0  ||  0  ||  0.06  ||  0.01  ||  2.343  || '9056306' ||
 * 52 ||  34.78  ||  -18.82  ||  6  ||  0  ||  0  ||  0  ||  1.09  ||  0.08  ||  2.968  || '7842136' ||
 * 53 ||  34.58  ||  -29.5  ||  2.62  ||  0  ||  0  ||  0  ||  1.42  ||  0.07  ||  3.525  || '5921021' ||
 * 54 ||  34.51  ||  -28.57  ||  2  ||  0  ||  0  ||  0  ||  0.12  ||  0.19  ||  1.081  || '7115698' ||
 * 55 ||  34.47  ||  -30.63  ||  2.21  ||  0  ||  0  ||  0  ||  1.57  ||  0.18  ||  4.038  || '7957875' ||
 * 56 ||  34.37  ||  -31.98  ||  0.99  ||  0  ||  0  ||  0  ||  0.34  ||  0.05  ||  3.553  || '7493042' ||
 * 57 ||  34.29  ||  -26.39  ||  3.01  ||  0  ||  0  ||  0  ||  0.58  ||  0.02  ||  1.476  || '5135469' ||
 * 58 ||  34.04  ||  -32.52  ||  1.04  ||  0  ||  0  ||  0  ||  0.81  ||  0.02  ||  3.1  || '7963108' ||
 * 59 ||  34.02  ||  -28.8  ||  1.95  ||  0  ||  0  ||  0  ||  0.34  ||  0.02  ||  2.642  || '7684808' ||
 * 60 ||  34  ||  -31.76  ||  1.56  ||  0  ||  0  ||  0.15  ||  2.15  ||  2  ||  4.592  || '7965868' ||
 * 61 ||  33.95  ||  -26.18  ||  2.97  ||  0  ||  0  ||  0  ||  0.57  ||  0  ||  3.255  || '5115021' ||
 * 62 ||  33.89  ||  -29.91  ||  1.61  ||  0  ||  0  ||  0  ||  0.46  ||  0.06  ||  3.132  || '7723709' ||
 * 63 ||  33.78  ||  -31.03  ||  1.9  ||  0  ||  0  ||  0  ||  1.49  ||  0.03  ||  2.368  || '5807003' ||
 * 64 ||  33.76  ||  -24.29  ||  3.39  ||  0  ||  0  ||  0  ||  0.37  ||  0.04  ||  2.433  || '5101823' ||
 * 65 ||  33.74  ||  -28.76  ||  1.86  ||  0  ||  0  ||  0  ||  0.3  ||  0.01  ||  2.93  || '5727879' ||
 * 66 ||  33.7  ||  -30.61  ||  0.9  ||  0.88  ||  0  ||  0  ||  1.18  ||  0.13  ||  4.048  || '7988034' ||
 * 67 ||  33.68  ||  -25.82  ||  3.22  ||  0  ||  0  ||  0  ||  1.05  ||  0.02  ||  3.306  || '7759524' ||
 * 68 ||  33.65  ||  -34.64  ||  0.54  ||  0  ||  0  ||  0  ||  1.42  ||  0.25  ||  3.934  || '7474459' ||
 * 69 ||  33.56  ||  -28.03  ||  2.63  ||  0  ||  0  ||  1.91  ||  0.41  ||  0.36  ||  2.679  || '6215486' ||
 * 70 ||  33.5  ||  -30.32  ||  1.98  ||  0  ||  0  ||  0  ||  2.55  ||  2.35  ||  3.778  || '5105504' ||
 * 71 ||  33.38  ||  -31.36  ||  0.75  ||  0  ||  0  ||  0  ||  0.13  ||  0.05  ||  1.859  || '5366906' ||
 * 72 ||  33.33  ||  -29.63  ||  1.65  ||  0  ||  0  ||  0  ||  1.68  ||  2.11  ||  3.437  || '9062377' ||
 * 73 ||  33.26  ||  -30.05  ||  1.5  ||  0  ||  0  ||  0  ||  1.83  ||  1.99  ||  4.788  || '9054741' ||
 * 74 ||  33.21  ||  -30.51  ||  1  ||  0  ||  0  ||  0  ||  0.16  ||  0  ||  1.465  || '9015782' ||
 * 75 ||  33.15  ||  -26.71  ||  1.6  ||  0.88  ||  0  ||  0  ||  0.53  ||  0.01  ||  2.9  || '7611472' ||
 * 76 ||  33.08  ||  -11.54  ||  7.73  ||  0  ||  0  ||  0  ||  0.94  ||  0.04  ||  3.171  || '5175515' ||
 * 77 ||  33  ||  -26.02  ||  2.82  ||  0  ||  0  ||  0  ||  0.75  ||  0  ||  1.875  || '7745372' ||
 * 78 ||  32.88  ||  -30.29  ||  0.98  ||  0  ||  0  ||  0  ||  0.18  ||  0.03  ||  2.534  || '9042832' ||
 * 79 ||  32.82  ||  -26.44  ||  2.34  ||  0  ||  0  ||  0  ||  0.58  ||  0.41  ||  2.187  || '5375243' ||
 * 80 ||  32.77  ||  -30.73  ||  0.88  ||  0  ||  0  ||  0.5  ||  0.24  ||  0.38  ||  2.442  || '7661649' ||
 * 81 ||  32.6  ||  -25.46  ||  2.82  ||  0  ||  0  ||  0  ||  0.84  ||  0.35  ||  3.013  || '7819337' ||
 * 82 ||  32.57  ||  -22.48  ||  3.84  ||  0  ||  0  ||  0  ||  0.82  ||  0.09  ||  4.052  || '6407567' ||
 * 83 ||  32.55  ||  -27.48  ||  1.93  ||  0  ||  0  ||  0  ||  0.39  ||  0.04  ||  3.843  || '5325354' ||
 * 84 ||  32.52  ||  -28.04  ||  2.12  ||  0  ||  0  ||  0.9  ||  0.8  ||  0.6  ||  4.514  || 9049724' ||
 * 85 ||  32.38  ||  -30.96  ||  1  ||  0  ||  0  ||  0  ||  0.81  ||  0.01  ||  3.156  || '5321784' ||
 * 86 ||  32.37  ||  -26.34  ||  2.68  ||  0  ||  0  ||  0  ||  1.04  ||  0.06  ||  1.89  || '7589277' ||
 * 87 ||  32.19  ||  -21.15  ||  3.76  ||  0  ||  0  ||  0  ||  0.17  ||  0.11  ||  4.017  || '7676310' ||
 * 88 ||  32.1  ||  -17.08  ||  5.87  ||  0  ||  0  ||  0  ||  1.31  ||  0.03  ||  3.462  || '5490779' ||
 * 89 ||  32.05  ||  -25.12  ||  2.34  ||  0  ||  0  ||  0  ||  0.05  ||  0.01  ||  1.984  || '7368624' ||
 * 90 ||  32.02  ||  -29.03  ||  1.55  ||  0  ||  0  ||  0  ||  1.24  ||  0.63  ||  2.539  || '9040674' ||
 * 91 ||  32  ||  -25.31  ||  3  ||  0  ||  0  ||  0  ||  1.22  ||  0.11  ||  3.901  || '6565654' ||
 * 92 ||  31.96  ||  -27.43  ||  2.06  ||  0  ||  0  ||  0  ||  0.87  ||  0.05  ||  3.885  || '5655793' ||
 * 93 ||  31.94  ||  -30.61  ||  1.46  ||  0  ||  0  ||  0  ||  1.54  ||  0.03  ||  3.137  || '7659306' ||
 * 94 ||  31.88  ||  -27.23  ||  1.99  ||  0  ||  0  ||  0  ||  0.75  ||  0.02  ||  2.52  || '5107288' ||
 * 95 ||  31.77  ||  -26.24  ||  1.97  ||  0  ||  0  ||  0  ||  0.2  ||  0.01  ||  2.262  || '6430967' ||
 * 96 ||  31.74  ||  -23.49  ||  2.99  ||  0  ||  0  ||  0  ||  0.37  ||  0.01  ||  3.624  || '7679150' ||
 * 97 ||  31.71  ||  -24.55  ||  2.99  ||  0  ||  0  ||  0  ||  0.93  ||  0.02  ||  2.831  || '6431688' ||
 * 98 ||  31.71  ||  -26.73  ||  1  ||  1  ||  0  ||  0  ||  0.53  ||  0.05  ||  2.556  || '7780474' ||
 * 99 ||  31.56  ||  -27.46  ||  1.69  ||  0  ||  0  ||  0  ||  0.56  ||  0.13  ||  2.668  || '5113142' ||
 * 100 ||  31.42  ||  -28.9  ||  1.26  ||  0  ||  0  ||  0  ||  0.7  ||  0.13  ||  3.201  || '5274837' ||
 * 101 ||  31.36  ||  -31.99  ||  0.54  ||  0  ||  0  ||  0  ||  1.13  ||  0.01  ||  3.221  || '5318411' ||
 * 102 ||  31.34  ||  -22.77  ||  3.42  ||  0  ||  0  ||  0  ||  1.2  ||  0.24  ||  1.377  || '5740453' ||
 * 103 ||  31.32  ||  -25.59  ||  2.52  ||  0  ||  0  ||  0  ||  0.93  ||  0  ||  1.812  || '9032552' ||
 * 104 ||  31.26  ||  -28.4  ||  1.03  ||  0.7  ||  0  ||  0  ||  1.19  ||  0.05  ||  3.457  || '7906118' ||
 * 105 ||  31.22  ||  -28.91  ||  1  ||  0  ||  0  ||  0  ||  0.35  ||  0.01  ||  2.112  || '5521475' ||
 * 106 ||  31.2  ||  -22.55  ||  2.93  ||  0  ||  0  ||  0  ||  0.08  ||  0.02  ||  3.707  || '7591231' ||
 * 107 ||  31.14  ||  -25.39  ||  2.31  ||  0  ||  0  ||  0  ||  0.91  ||  0.65  ||  3.092  || '7539979' ||
 * 108 ||  30.86  ||  -26.03  ||  2.66  ||  0  ||  0  ||  0  ||  1.59  ||  0.01  ||  3.096  || '5521698' ||
 * 109 ||  30.84  ||  -29.63  ||  0.79  ||  0  ||  0  ||  0  ||  1.97  ||  2.77  ||  3.504  || '7383117' ||
 * 110 ||  30.84  ||  -23.64  ||  2.99  ||  0  ||  0  ||  0  ||  0.91  ||  0.05  ||  2.856  || '7668608' ||
 * 111 ||  30.84  ||  -25  ||  2.3  ||  0  ||  0  ||  0.14  ||  1.51  ||  2.07  ||  3.714  || '7965658' ||
 * 112 ||  30.84  ||  -24.41  ||  1.35  ||  1.48  ||  0  ||  0  ||  1.07  ||  0.04  ||  3.29  || '7971092' ||
 * 113 ||  30.78  ||  -28.05  ||  1.2  ||  0  ||  0  ||  0  ||  0.51  ||  0.15  ||  3.707  || '7726200' ||
 * 114 ||  30.78  ||  -26.42  ||  1.86  ||  0  ||  0  ||  0  ||  1.29  ||  1.35  ||  3.136  || '7740160' ||
 * 115 ||  30.77  ||  -28.34  ||  1.38  ||  0  ||  0  ||  0  ||  0.86  ||  0.01  ||  2.063  || '6050973' ||
 * 116 ||  30.72  ||  -26.19  ||  1.51  ||  0  ||  0  ||  0  ||  0.01  ||  0  ||  0.477  || '7674251' ||
 * 117 ||  30.68  ||  -16.84  ||  4.95  ||  0  ||  0  ||  0  ||  0.52  ||  0.01  ||  3.028  || '5490061' ||
 * 118 ||  30.67  ||  -31.91  ||  0  ||  0  ||  0  ||  0  ||  0.74  ||  0.23  ||  4.181  || '7743670' ||
 * 119 ||  30.62  ||  -24.19  ||  2.63  ||  0  ||  0  ||  0  ||  1.79  ||  2.11  ||  3.082  || '7964554' ||
 * 120 ||  30.55  ||  -29.15  ||  1.22  ||  0  ||  0  ||  0  ||  1.18  ||  0.1  ||  3.211  || '5404845' ||
 * 121 ||  30.51  ||  -24.67  ||  2  ||  0  ||  0  ||  0  ||  0.09  ||  0.02  ||  1.198  || '9043424' ||
 * 122 ||  30.48  ||  -26.46  ||  1.91  ||  0  ||  0  ||  0  ||  0.87  ||  0.03  ||  3.023  || '9037367' ||
 * 123 ||  30.33  ||  -22.02  ||  2.83  ||  0  ||  0  ||  0  ||  0.1  ||  0.01  ||  2.33  || '5324761' ||
 * 124 ||  30.33  ||  -27.54  ||  1.47  ||  0  ||  0  ||  0  ||  0.82  ||  0.03  ||  2.624  || '5486215' ||
 * 125 ||  30.24  ||  -23.18  ||  2.77  ||  0  ||  0  ||  0  ||  0.63  ||  0.01  ||  2.575  || '7731013' ||
 * 126 ||  30.23  ||  -28.28  ||  1  ||  0  ||  0  ||  0  ||  0.88  ||  0.65  ||  3.014  || '5256974' ||
 * 127 ||  30.22  ||  -27.3  ||  1.99  ||  0  ||  0  ||  0  ||  1.55  ||  0.01  ||  2.25  || '5573924' ||
 * 128 ||  30.18  ||  -29.27  ||  0.77  ||  0  ||  0  ||  0  ||  0.73  ||  0.06  ||  3.477  || '7958373' ||
 * 129 ||  30.08  ||  -30.17  ||  0  ||  0  ||  0  ||  0.04  ||  0.48  ||  0.79  ||  2.569  || '5241813' ||
 * 130 ||  30.07  ||  -26.02  ||  1.61  ||  0  ||  0  ||  0  ||  0.39  ||  0  ||  2.311  || '9020863' ||
 * 131 ||  30.04  ||  -29.65  ||  0.8  ||  0  ||  0  ||  0  ||  1.04  ||  0.06  ||  3.877  || '7748043' ||
 * 132 ||  30.02  ||  -27.53  ||  1  ||  0  ||  0  ||  0  ||  0.26  ||  0.02  ||  2.728  || '9025155' ||
 * 133 ||  29.9  ||  -22.63  ||  2.98  ||  0  ||  0  ||  0  ||  0.84  ||  0.02  ||  3.296  || '7903592' ||
 * 134 ||  29.85  ||  -33.31  ||  0.9  ||  0  ||  0  ||  0  ||  3.37  ||  0.19  ||  4.268  || '5784214' ||
 * 135 ||  29.85  ||  -28.12  ||  0.77  ||  0  ||  0  ||  0  ||  0.29  ||  0  ||  2.232  || '7651662' ||
 * 136 ||  29.84  ||  -23.16  ||  2.54  ||  0  ||  0  ||  0  ||  0.48  ||  0.02  ||  2.208  || '7992668' ||
 * 137 ||  29.8  ||  -27.74  ||  1  ||  0  ||  0  ||  0  ||  0.49  ||  0  ||  2.444  || '7987751' ||
 * 138 ||  29.79  ||  -23.45  ||  2.41  ||  0  ||  0  ||  0  ||  0.44  ||  0  ||  0.98  || '5320628' ||
 * 139 ||  29.72  ||  -25.49  ||  1.78  ||  0  ||  0  ||  0  ||  0.79  ||  0.3  ||  4.04  || '5308420' ||
 * 140 ||  29.72  ||  -24.8  ||  1  ||  0.9  ||  0  ||  0  ||  0.46  ||  0.16  ||  4.393  || '7976083' ||
 * 141 ||  29.71  ||  -28.47  ||  0.98  ||  0  ||  0  ||  0  ||  0.85  ||  0.02  ||  3.169  || '6352243' ||
 * 142 ||  29.69  ||  -25.55  ||  1.54  ||  0  ||  0  ||  0  ||  0.25  ||  0.01  ||  2.324  || '7739941' ||
 * 143 ||  29.59  ||  -22.82  ||  2.82  ||  0  ||  0  ||  0  ||  0.86  ||  0.03  ||  2.469  || '5308586' ||
 * 144 ||  29.55  ||  -14.57  ||  5.31  ||  0  ||  0  ||  0  ||  0.48  ||  0.01  ||  1.791  || '5731094' ||
 * 145 ||  29.52  ||  -27.83  ||  1.02  ||  0  ||  0  ||  0  ||  0.69  ||  0.01  ||  2.166  || '7642712' ||
 * 146 ||  29.49  ||  -27.6  ||  1.8  ||  0  ||  0  ||  0  ||  1.76  ||  0.02  ||  0.93  || '5245534' ||
 * 147 ||  29.48  ||  -25.21  ||  2.25  ||  0  ||  0  ||  2.29  ||  1.62  ||  3.04  ||  2.26  || '9049272' ||
 * 148 ||  29.45  ||  -23.94  ||  2.39  ||  0  ||  0  ||  0  ||  1.06  ||  0.22  ||  4.185  || '7683213' ||
 * 149 ||  29.43  ||  -27.15  ||  1.23  ||  0  ||  0  ||  0  ||  0.76  ||  0.11  ||  3.004  || '5227340' ||
 * 150 ||  29.42  ||  -26.77  ||  0.93  ||  0  ||  0  ||  0  ||  0.1  ||  0.06  ||  3.005  || '5684393' ||
 * 151 ||  29.37  ||  -23.45  ||  0.98  ||  1  ||  0  ||  0  ||  0  ||  0  ||  1.138  || '7748819' ||
 * 152 ||  29.31  ||  -24.07  ||  1.78  ||  0  ||  0  ||  0  ||  1.13  ||  2.11  ||  3.325  || '7959170' ||
 * 153 ||  29.3  ||  -24.96  ||  1.56  ||  0  ||  0  ||  0  ||  0.17  ||  0.01  ||  2.066  || '5373349' ||
 * 154 ||  29.27  ||  -27.99  ||  1  ||  0  ||  0  ||  0  ||  0.89  ||  0.05  ||  2.766  || '5402310' ||
 * 155 ||  29.25  ||  -27.51  ||  0.72  ||  0  ||  0  ||  0  ||  0.22  ||  0.02  ||  2.367  || '7747481' ||
 * 156 ||  29.21  ||  -25.5  ||  1.52  ||  0  ||  0  ||  0  ||  0.52  ||  0.07  ||  3.227  || '6540387' ||
 * 157 ||  29.19  ||  -27.99  ||  0.54  ||  0  ||  0  ||  0  ||  1.33  ||  2.23  ||  3.378  || '7757183' ||
 * 158 ||  29.16  ||  -21.82  ||  2.86  ||  0  ||  0  ||  0  ||  0.63  ||  0  ||  2.542  || '7814264' ||
 * 159 ||  29.15  ||  -15.12  ||  4  ||  1  ||  0  ||  0  ||  0.5  ||  0.04  ||  1.77  || '7397377' ||
 * 160 ||  29.15  ||  -26.57  ||  1  ||  0  ||  0  ||  0  ||  0.25  ||  0.03  ||  2.86  || '7692562' ||
 * 161 ||  29.12  ||  -28.03  ||  1.55  ||  0  ||  0  ||  0  ||  1.97  ||  0.36  ||  4.068  || '7855467' ||
 * 162 ||  29.09  ||  -20.54  ||  2.98  ||  0  ||  0  ||  0  ||  0.32  ||  0.24  ||  1.636  || '5316636' ||
 * 163 ||  29.06  ||  -24.59  ||  1.91  ||  0  ||  0  ||  0  ||  0.69  ||  0.13  ||  2.362  || '7727707' ||
 * 164 ||  29.05  ||  -26.24  ||  0.59  ||  0.77  ||  0  ||  0  ||  0.65  ||  0.02  ||  2.83  || '7516821' ||
 * 165 ||  29.02  ||  -26.43  ||  1.51  ||  0  ||  0  ||  0  ||  1  ||  0.01  ||  3.449  || '7650920' ||
 * 166 ||  29.02  ||  -26.47  ||  1  ||  0  ||  0  ||  0  ||  0.23  ||  0  ||  2.678  || '7926747' ||
 * 167 ||  28.93  ||  -21.66  ||  2.74  ||  0  ||  0  ||  0  ||  0.48  ||  0.02  ||  2.117  || '5672721' ||
 * 168 ||  28.85  ||  -29.03  ||  0.26  ||  0  ||  0  ||  0  ||  1.28  ||  1.6  ||  1.909  || '5130645' ||
 * 169 ||  28.79  ||  -18.42  ||  3.49  ||  0  ||  0  ||  0  ||  0.05  ||  0.01  ||  1.471  || '7278181' ||
 * 170 ||  28.79  ||  -28.88  ||  0.32  ||  0  ||  0  ||  0  ||  1.45  ||  1.85  ||  2.73  || '9050714' ||
 * 171 ||  28.72  ||  -15.12  ||  4.55  ||  0  ||  0  ||  0  ||  0.04  ||  0.02  ||  2.235  || '7907769' ||
 * 172 ||  28.53  ||  -23.8  ||  1.96  ||  0  ||  0  ||  0  ||  1.63  ||  2.12  ||  2.563  || '5230085' ||
 * 173 ||  28.48  ||  -20.23  ||  3.36  ||  0  ||  0  ||  0  ||  0.96  ||  0.03  ||  2.11  || '9049267' ||
 * 174 ||  28.43  ||  -23.01  ||  2.9  ||  0  ||  0  ||  0  ||  1.7  ||  0.09  ||  3.621  || '6434273' ||
 * 175 ||  28.36  ||  -23.46  ||  1.74  ||  0  ||  0  ||  0  ||  0.18  ||  0.05  ||  1.531  || '7635462' ||
 * 176 ||  28.35  ||  -21.64  ||  2.91  ||  0  ||  0  ||  0  ||  3.17  ||  4.32  ||  3.587  || '7541177' ||
 * 177 ||  28.28  ||  -25.09  ||  1.31  ||  0  ||  0  ||  0  ||  0.52  ||  0.15  ||  3.78  || '7722615' ||
 * 178 ||  28.26  ||  -15.05  ||  4.63  ||  0  ||  0  ||  0  ||  1.69  ||  2.71  ||  2.12  || '9011332' ||
 * 179 ||  28.2  ||  -23.44  ||  1.93  ||  0  ||  0  ||  0  ||  0.52  ||  0.02  ||  2.783  || '9016219' ||
 * 180 ||  28.2  ||  -25.45  ||  1.5  ||  0  ||  0  ||  0  ||  0.87  ||  0  ||  0.554  || '9041798' ||
 * 181 ||  28.16  ||  -26.12  ||  1  ||  0  ||  0  ||  0  ||  0.58  ||  0.2  ||  3.697  || '7828364' ||
 * 182 ||  28.14  ||  -22.98  ||  1.84  ||  0  ||  0  ||  0  ||  1.22  ||  2.08  ||  3.562  || '7825083' ||
 * 183 ||  28.1  ||  -26.33  ||  0.62  ||  0  ||  0  ||  0  ||  0.05  ||  0  ||  1.874  || '9021968' ||
 * 184 ||  27.98  ||  -24.54  ||  1.47  ||  0  ||  0  ||  0  ||  0.52  ||  0.08  ||  2.843  || '6432007' ||
 * 185 ||  27.97  ||  -19.2  ||  2.94  ||  0  ||  0  ||  0  ||  0.04  ||  0.02  ||  2.751  || '7687650' ||
 * 186 ||  27.94  ||  -21.93  ||  2.96  ||  0  ||  0  ||  0  ||  1.66  ||  0.46  ||  3.026  || '7893858' ||
 * 187 ||  27.88  ||  -22.95  ||  2.28  ||  0  ||  0  ||  0  ||  0.96  ||  0  ||  2.191  || '9039578' ||
 * 188 ||  27.83  ||  -26.83  ||  0.38  ||  0  ||  0  ||  0  ||  0.08  ||  0  ||  2.622  || '7690618' ||
 * 189 ||  27.7  ||  -25.3  ||  0.99  ||  0  ||  0  ||  0  ||  0.29  ||  0.01  ||  0.984  || '5363541' ||
 * 190 ||  27.69  ||  -20.02  ||  3  ||  0  ||  0  ||  0  ||  1.08  ||  0.82  ||  1.069  || '7734200' ||
 * 191 ||  27.65  ||  -24.49  ||  1.53  ||  0  ||  0  ||  0  ||  0.73  ||  0.02  ||  2.582  || '7682152' ||
 * 192 ||  27.64  ||  -22.86  ||  1.74  ||  0  ||  0  ||  0  ||  0.22  ||  0  ||  2.639  || '5787786' ||
 * 193 ||  27.59  ||  -22.49  ||  1.97  ||  0  ||  0  ||  0  ||  0.44  ||  0.06  ||  1.555  || '5310038' ||
 * 194 ||  27.57  ||  -17.9  ||  3.55  ||  0  ||  0  ||  0  ||  0.52  ||  0.06  ||  3.006  || '9031046' ||
 * 195 ||  27.56  ||  -19.76  ||  3  ||  0  ||  0  ||  0  ||  0.67  ||  0.13  ||  4.071  || '7848095' ||
 * 196 ||  27.55  ||  -19.18  ||  3  ||  0  ||  0  ||  0  ||  0.67  ||  0.42  ||  4.445  || '7491682' ||
 * 197 ||  27.55  ||  -18.65  ||  3.75  ||  0  ||  0  ||  0  ||  1.27  ||  0.08  ||  3.613  || '7663126' ||
 * 198 ||  27.43  ||  -23.3  ||  1.64  ||  0  ||  0  ||  0  ||  0.41  ||  0.04  ||  3.173  || '9022356' ||
 * 199 ||  27.41  ||  -28.68  ||  1  ||  0  ||  0  ||  0  ||  2.24  ||  0.22  ||  3.185  || '5655896' ||
 * 200 ||  27.39  ||  -23.31  ||  1.7  ||  0  ||  0  ||  0  ||  0.53  ||  0.02  ||  3.131  || '7777802' ||
 * 201 ||  27.37  ||  -22.46  ||  1.95  ||  0  ||  0  ||  0  ||  0.49  ||  0.01  ||  0.731  || '9020176' ||
 * 202 ||  27.34  ||  -18.54  ||  2.98  ||  0  ||  0  ||  0  ||  0.1  ||  0.04  ||  0.968  || '6017107' ||
 * 203 ||  27.31  ||  -20.42  ||  2.45  ||  0  ||  0  ||  0  ||  0.23  ||  0  ||  1.228  || '5380171' ||
 * 204 ||  27.28  ||  -19.13  ||  1.92  ||  0.88  ||  0  ||  0  ||  1.23  ||  2.23  ||  2.821  || '7965915' ||
 * 205 ||  27.27  ||  -22.45  ||  1.81  ||  0  ||  0  ||  0  ||  0.31  ||  0.02  ||  2.492  || '5623003' ||
 * 206 ||  27.25  ||  -26.57  ||  1.23  ||  0  ||  0  ||  0  ||  1.53  ||  0.04  ||  2.988  || '5102939' ||
 * 207 ||  27.21  ||  -19.46  ||  2.85  ||  0  ||  0  ||  0  ||  0.44  ||  0.02  ||  2.658  || '7686962' ||
 * 208 ||  27.21  ||  -28.03  ||  0  ||  0  ||  0  ||  0  ||  0.41  ||  0  ||  2.384  || '9003877' ||
 * 209 ||  27.2  ||  -22.53  ||  1.8  ||  0  ||  0  ||  0  ||  0.37  ||  0.01  ||  2.724  || '7934500' ||
 * 210 ||  27.18  ||  -22.96  ||  1.78  ||  0  ||  0  ||  0  ||  0.58  ||  0.02  ||  2.55  || '7761278' ||
 * 211 ||  27.16  ||  -20.89  ||  2.96  ||  0  ||  0  ||  0  ||  1.31  ||  0  ||  2.39  || '5113066' ||
 * 212 ||  27.11  ||  -23.71  ||  2.28  ||  0  ||  0  ||  0  ||  1.74  ||  0.02  ||  1.948  || '9052908' ||
 * 213 ||  27.09  ||  -22.82  ||  1.45  ||  0  ||  0  ||  0  ||  0.04  ||  0.01  ||  1.576  || '9029346' ||
 * 214 ||  27.07  ||  -17.35  ||  4  ||  0  ||  0  ||  0  ||  2.34  ||  2.4  ||  2.944  || '5531766' ||
 * 215 ||  27.05  ||  -25.53  ||  0.8  ||  0  ||  0  ||  0  ||  0.45  ||  0  ||  1.88  || '5307981' ||
 * 216 ||  27.05  ||  -22.02  ||  1.95  ||  0  ||  0  ||  0  ||  0.41  ||  0  ||  1.788  || '7530226' ||
 * 217 ||  27.02  ||  -27.04  ||  0.62  ||  0  ||  0  ||  0  ||  1.28  ||  0.61  ||  2.859  || '9024121' ||
 * 218 ||  26.97  ||  -18.5  ||  2.88  ||  0  ||  0  ||  0  ||  0.1  ||  0.02  ||  1.003  || '5306063' ||
 * 219 ||  26.92  ||  -29.33  ||  0.03  ||  0  ||  0  ||  0  ||  1.32  ||  0.11  ||  3.33  || '7954091' ||
 * 220 ||  26.85  ||  -17.73  ||  3.22  ||  0  ||  0  ||  0  ||  0.31  ||  0.06  ||  2.796  || '7928256' ||
 * 221 ||  26.62  ||  -21.17  ||  2.35  ||  0  ||  0  ||  0  ||  1.09  ||  0.56  ||  3.312  || '7886522' ||
 * 222 ||  26.59  ||  -22.09  ||  1.67  ||  0  ||  0  ||  0  ||  0.26  ||  0.01  ||  2.123  || '5483223' ||
 * 223 ||  26.55  ||  -21.26  ||  1.9  ||  0  ||  0  ||  0  ||  0.2  ||  0  ||  1.57  || '4102009' ||
 * 224 ||  26.54  ||  -22.9  ||  1.82  ||  0  ||  0  ||  0  ||  0.93  ||  0.04  ||  1.718  || '7800400' ||
 * 225 ||  26.48  ||  -20.57  ||  2.79  ||  0  ||  0  ||  0  ||  1.3  ||  0.16  ||  3.532  || '5552937' ||
 * 226 ||  26.41  ||  -21.85  ||  2.1  ||  0  ||  0  ||  0  ||  0.89  ||  0.03  ||  3.407  || '5784281' ||
 * 227 ||  26.37  ||  -24.25  ||  1  ||  0  ||  0  ||  0  ||  0.45  ||  0.03  ||  3.103  || '7752611' ||
 * 228 ||  26.31  ||  -24.24  ||  1  ||  0  ||  0  ||  0  ||  1.4  ||  1.86  ||  0.695  || '7634187' ||
 * 229 ||  26.3  ||  -20.91  ||  2.17  ||  0  ||  0  ||  0  ||  1.84  ||  2.56  ||  1.939  || '7680523' ||
 * 230 ||  26.29  ||  -24.28  ||  1.13  ||  0  ||  0  ||  0  ||  1.64  ||  1.91  ||  2.67  || '7736742' ||
 * 231 ||  26.26  ||  -23.22  ||  1.44  ||  0  ||  0  ||  0  ||  0.8  ||  0.32  ||  4.723  || '7869434' ||
 * 232 ||  26.19  ||  -28.28  ||  0  ||  0  ||  0  ||  0  ||  1.05  ||  0.01  ||  2.754  || '7116397' ||
 * 233 ||  26.15  ||  -26.8  ||  0.55  ||  0  ||  0  ||  0  ||  1.15  ||  0.01  ||  2.635  || '5558102' ||
 * 234 ||  26.09  ||  -24.7  ||  1  ||  0  ||  0  ||  0  ||  0.82  ||  0.02  ||  2.14  || '7674411' ||
 * 235 ||  26.04  ||  -21.24  ||  1.6  ||  0  ||  0  ||  0  ||  0.02  ||  0.03  ||  1.902  || '5225638' ||
 * 236 ||  25.68  ||  -23.22  ||  0.93  ||  0  ||  0  ||  0  ||  0.17  ||  0.01  ||  1.426  || '5314675' ||
 * 237 ||  25.58  ||  -25.04  ||  0.5  ||  0  ||  0  ||  0  ||  0.49  ||  0.04  ||  3.527  || '7630579' ||
 * 238 ||  25.36  ||  -22.71  ||  1  ||  0  ||  0  ||  0  ||  0.17  ||  0  ||  1.366  || '5939731' ||
 * 239 ||  25.31  ||  -21.11  ||  1.7  ||  0  ||  0  ||  0  ||  0.45  ||  0.01  ||  2.27  || '7745425' ||
 * 240 ||  25.3  ||  -22.48  ||  1  ||  0  ||  0  ||  0  ||  0.26  ||  0.24  ||  2.717  || '7854051' ||
 * 241 ||  25.26  ||  -24.16  ||  0.37  ||  0  ||  0  ||  0  ||  0  ||  0  ||  1.06  || '6755121' ||
 * 242 ||  25.22  ||  -22.21  ||  1.58  ||  0  ||  0  ||  0  ||  0.89  ||  0.05  ||  2.681  || '5252659' ||
 * 243 ||  25.03  ||  -24.42  ||  0.62  ||  0  ||  0  ||  0  ||  0.68  ||  0.07  ||  3.441  || '7690200' ||
 * 244 ||  24.83  ||  -19.23  ||  2  ||  0  ||  0  ||  0  ||  0.2  ||  0  ||  2.244  || '9018640' ||
 * 245 ||  24.78  ||  -27.29  ||  0.03  ||  0  ||  0  ||  0  ||  2.07  ||  1.54  ||  2.538  || '7795974' ||
 * 246 ||  24.74  ||  -19.84  ||  1.74  ||  0  ||  0  ||  0  ||  0.15  ||  0  ||  1.369  || '5614186' ||
 * 247 ||  24.66  ||  -22.01  ||  0.94  ||  0  ||  0  ||  0  ||  0.1  ||  0.02  ||  0.521  || '7934371' ||
 * 248 ||  24.58  ||  -25.98  ||  0  ||  0  ||  0  ||  0  ||  0.71  ||  0.01  ||  2.055  || '7777514' ||
 * 249 ||  24.52  ||  -21.72  ||  1  ||  0  ||  0  ||  0  ||  0.12  ||  0.05  ||  2.724  || '7733216' ||
 * 250 ||  24.48  ||  -25.57  ||  0  ||  0  ||  0  ||  0  ||  0.7  ||  0.02  ||  2.933  || '7600192' ||
 * 251 ||  24.43  ||  -24.78  ||  0  ||  0  ||  0  ||  0  ||  0.19  ||  0.03  ||  2.101  || '5325654' ||
 * 252 ||  24.4  ||  -14.3  ||  3.88  ||  0  ||  0  ||  0  ||  1.17  ||  0.79  ||  0.668  || '9031574' ||
 * 253 ||  24.37  ||  -18.8  ||  2  ||  0  ||  0  ||  0  ||  0.22  ||  0  ||  1.232  || '5666147' ||
 * 254 ||  24.35  ||  -20.19  ||  1.56  ||  0  ||  0  ||  0  ||  0.27  ||  0.02  ||  2.359  || '5253555' ||
 * 255 ||  24.26  ||  -25.8  ||  0.64  ||  0  ||  0  ||  1.35  ||  2  ||  1.88  ||  3.169  || '7495305' ||
 * 256 ||  24.22  ||  -22.84  ||  0.62  ||  0  ||  0  ||  0  ||  0.51  ||  0.53  ||  1.215  || '7662712' ||
 * 257 ||  24.2  ||  -16.41  ||  3.12  ||  0  ||  0  ||  0  ||  2.38  ||  3.18  ||  1.858  || '5319694' ||
 * 258 ||  24.18  ||  -19.58  ||  1.63  ||  0  ||  0  ||  0  ||  0.16  ||  0.02  ||  2.095  || '6045092' ||
 * 259 ||  24.17  ||  -15.69  ||  2.85  ||  0  ||  0  ||  0  ||  0.07  ||  0.05  ||  0.52  || '5175423' ||
 * 260 ||  24.07  ||  -21.74  ||  1  ||  0  ||  0  ||  0  ||  0.33  ||  0.01  ||  1.034  || '5728158' ||
 * 261 ||  24  ||  -24.31  ||  0.19  ||  0  ||  0  ||  0  ||  0.45  ||  0.01  ||  2.619  || '5131604' ||
 * 262 ||  23.95  ||  -25.28  ||  0  ||  0  ||  0  ||  0  ||  0.76  ||  0.18  ||  0.978  || '7637949' ||
 * 263 ||  23.92  ||  -18.82  ||  2  ||  0  ||  0  ||  0  ||  0.62  ||  0.33  ||  1.327  || '5225540' ||
 * 264 ||  23.74  ||  -23.98  ||  0.8  ||  0  ||  0  ||  0.59  ||  1.1  ||  0.04  ||  2.987  || '7952016' ||
 * 265 ||  23.6  ||  -22.76  ||  0.58  ||  0  ||  0  ||  0  ||  1.51  ||  2.14  ||  2.229  || '7955386' ||
 * 266 ||  23.59  ||  -20.49  ||  1.5  ||  0  ||  0  ||  0  ||  0.72  ||  0.02  ||  2.13  || '5737089' ||
 * 267 ||  23.57  ||  -15.05  ||  2.98  ||  0  ||  0  ||  0  ||  0.23  ||  0.06  ||  4.215  || '7984791' ||
 * 268 ||  23.43  ||  -23.47  ||  0  ||  0  ||  0  ||  0  ||  0.02  ||  0.01  ||  3.118  || '5304628' ||
 * 269 ||  23.24  ||  -21.42  ||  0.64  ||  0  ||  0  ||  0  ||  0.05  ||  0  ||  0.531  || '4022614' ||
 * 270 ||  23.23  ||  -15.13  ||  2.8  ||  0  ||  0  ||  0  ||  0.15  ||  0  ||  1.656  || '7960310' ||
 * 271 ||  23.18  ||  -17.22  ||  2  ||  0  ||  0  ||  0  ||  0.04  ||  0.04  ||  0.652  || '5309703' ||
 * 272 ||  23.06  ||  -20.06  ||  1  ||  0  ||  0  ||  0  ||  0  ||  0  ||  1.18  || '5102936' ||
 * 273 ||  22.95  ||  -20.61  ||  0.98  ||  0  ||  0  ||  0  ||  0.29  ||  0  ||  1.882  || '9016814' ||
 * 274 ||  22.92  ||  -15.69  ||  2.58  ||  0  ||  0  ||  0  ||  0.25  ||  0  ||  2.203  || '9031350' ||
 * 275 ||  22.91  ||  -22.97  ||  0  ||  0  ||  0  ||  0.21  ||  0.04  ||  0.23  ||  2.189  || '7664184' ||
 * 276 ||  22.66  ||  -19.66  ||  1  ||  0  ||  0  ||  0  ||  0  ||  0  ||  0.813  || '5351178' ||
 * 277 ||  22.65  ||  -12.49  ||  3.72  ||  0  ||  0  ||  0.1  ||  0.87  ||  0.8  ||  2.554  || '5930362' ||
 * 278 ||  22.43  ||  -17.47  ||  1.82  ||  0  ||  0  ||  0  ||  0.87  ||  1.13  ||  2.362  || '5235377' ||
 * 279 ||  22.38  ||  -22.58  ||  0  ||  0  ||  0  ||  0  ||  0.1  ||  0  ||  1.968  || '5635486' ||
 * 280 ||  22.28  ||  -14.32  ||  2.7  ||  0  ||  0  ||  0  ||  0.08  ||  0  ||  1.308  || '5113883' ||
 * 281 ||  22.27  ||  -19.32  ||  1  ||  0  ||  0  ||  0  ||  0.48  ||  0.91  ||  1.119  || '5403251' ||
 * 282 ||  22.14  ||  -17.55  ||  1.93  ||  0  ||  0  ||  0  ||  0.69  ||  0.2  ||  2.096  || '5481947' ||
 * 283 ||  22.05  ||  -20.89  ||  0.69  ||  0  ||  0  ||  0  ||  0.46  ||  0.01  ||  2.017  || '5880585' ||
 * 284 ||  21.98  ||  -15.88  ||  2.5  ||  0  ||  0  ||  0  ||  0.71  ||  0.02  ||  0.615  || '9040725' ||
 * 285 ||  21.95  ||  -11.52  ||  3.97  ||  0  ||  0  ||  0  ||  0.78  ||  0.03  ||  2.634  || '7774207' ||
 * 286 ||  21.8  ||  -17.84  ||  1.96  ||  0  ||  0  ||  0  ||  1.78  ||  1.63  ||  2.403  || '7743596' ||
 * 287 ||  21.69  ||  -18.71  ||  1  ||  0  ||  0  ||  0  ||  0.01  ||  0  ||  1.463  || '9032389' ||
 * 288 ||  21.59  ||  -19.15  ||  0.99  ||  0  ||  0  ||  0  ||  0.27  ||  0  ||  1.085  || '7664088' ||
 * 289 ||  21.37  ||  -15.61  ||  2  ||  0  ||  0  ||  0  ||  0.12  ||  0  ||  1.926  || '4000001' ||
 * 290 ||  21.36  ||  -20.91  ||  1  ||  0.04  ||  0  ||  0  ||  1.35  ||  0.01  ||  1.908  || '5865114' ||
 * 291 ||  21.23  ||  -13.63  ||  3.34  ||  0  ||  0  ||  0  ||  1.23  ||  0.05  ||  0.577  || '5109432' ||
 * 292 ||  21.03  ||  -20.7  ||  0.88  ||  0  ||  0  ||  0.51  ||  1.97  ||  2.14  ||  3.249  || '5145823' ||
 * 293 ||  20.99  ||  -17.97  ||  1.02  ||  0  ||  0  ||  0  ||  0.02  ||  0  ||  2.099  || '5305260' ||
 * 294 ||  20.84  ||  -16.48  ||  1.46  ||  0  ||  0  ||  0  ||  0.23  ||  0.45  ||  1.783  || '7549075' ||
 * 295 ||  19.82  ||  -16.71  ||  1.23  ||  0  ||  0  ||  0  ||  0.29  ||  0  ||  0.604  || '9035977' ||
 * 296 ||  19.8  ||  -15.97  ||  1.62  ||  0  ||  0  ||  0  ||  0.6  ||  0.14  ||  3.032  || '5303377' ||
 * 297 ||  19.5  ||  -14.25  ||  2.58  ||  0  ||  0  ||  0  ||  1.24  ||  0  ||  1.166  || '9033391' ||
 * 298 ||  19.36  ||  -11.22  ||  3  ||  0  ||  0  ||  0  ||  0.48  ||  0.09  ||  2.85  || '9018385' ||
 * 299 ||  19.35  ||  -10.95  ||  3  ||  0  ||  0  ||  0  ||  0.3  ||  0.01  ||  0.996  || '4022605' ||
 * 300 ||  19.25  ||  -15.48  ||  1.3  ||  0  ||  0  ||  0  ||  0.21  ||  0.29  ||  0.36  || '9040279' ||
 * 301 ||  19.17  ||  -8.75  ||  3.55  ||  0  ||  0  ||  0  ||  0.12  ||  0.01  ||  0.449  || '4004234' ||
 * 302 ||  18.68  ||  -12.92  ||  1.99  ||  0  ||  0  ||  0  ||  0.11  ||  0  ||  1.23  || '5661991' ||
 * 303 ||  17.78  ||  -12.97  ||  1.72  ||  0  ||  0  ||  0  ||  0.4  ||  0.46  ||  1.233  || '7633686' ||
 * 304 ||  16.69  ||  -13.95  ||  1  ||  0  ||  0  ||  0  ||  0.13  ||  0.01  ||  1.394  || '9028071' ||
 * 305 ||  16.04  ||  -10.11  ||  1.99  ||  0  ||  0  ||  0  ||  1.51  ||  2.97  ||  0.739  || '9016568' ||
 * 306 ||  12.47  ||  -10.78  ||  0.6  ||  0  ||  0  ||  0  ||  0.05  ||  0.01  ||  0.583  || '9014885' ||

//10/28/13:// 3Q6X Docking with CB306 in GOLD
 * Table 2:** 3Q6Xdocked to CB306 in GOLD.


 * # || Score || S(PLP) || S(hbond) || S(cho) || S(metal) || DE(clash) || DE(tors) || intcor || time || name ||
 * 1 ||  97.74  ||  -41.74  ||  2.18  ||  0  ||  8.69  ||  0  ||  1.35  ||  0.01  ||  27.569  || '5175515' ||
 * 2 ||  95.79  ||  -53.1  ||  0.62  ||  0  ||  7.34  ||  0  ||  1.76  ||  0.31  ||  28.672  || '7726180' ||
 * 3 ||  95.13  ||  -55.98  ||  0  ||  0  ||  6.82  ||  0  ||  2.03  ||  2.19  ||  58.504  || '5531766' ||
 * 4 ||  94.32  ||  -53.57  ||  0  ||  0  ||  6.96  ||  0  ||  0.54  ||  0.06  ||  38.006  || '5282931' ||
 * 5 ||  93.19  ||  -40.31  ||  1.96  ||  0  ||  7.99  ||  0  ||  0.53  ||  0.11  ||  56.751  || '5192853' ||
 * 6 ||  91.99  ||  -76.57  ||  0.6  ||  0.96  ||  2  ||  0  ||  0.72  ||  0.11  ||  35.223  || '9027311' ||
 * 7 ||  91.84  ||  -51.6  ||  0  ||  0  ||  6.88  ||  0  ||  0.57  ||  0.08  ||  39.87  || '7575772' ||
 * 8 ||  89.58  ||  -60.17  ||  2.11  ||  0  ||  3.98  ||  0  ||  0.44  ||  0.03  ||  74.036  || '7746693' ||
 * 9 ||  87.9  ||  -39.14  ||  1  ||  0  ||  7.99  ||  0.98  ||  0.63  ||  0.06  ||  61.183  || '5467784' ||
 * 10 ||  87.48  ||  -43.36  ||  0  ||  0  ||  7.61  ||  0  ||  1.87  ||  2.22  ||  15.206  || '6629394' ||
 * 11 ||  87.03  ||  -44.38  ||  0.9  ||  0  ||  6.94  ||  0  ||  1.88  ||  2.09  ||  59.386  || '7676009' ||
 * 12 ||  86.37  ||  -43.54  ||  0.84  ||  0  ||  6.93  ||  0  ||  0.65  ||  0.02  ||  10.388  || '5101823' ||
 * 13 ||  85.35  ||  -41.94  ||  1.04  ||  0  ||  6.97  ||  0  ||  0.8  ||  0.06  ||  49.079  || '6407567' ||
 * 14 ||  84.7  ||  -85.67  ||  0  ||  0  ||  0  ||  0  ||  0.8  ||  0.62  ||  83.242  || '7558664' ||
 * 15 ||  83.42  ||  -53.2  ||  3.13  ||  0  ||  3.5  ||  0  ||  1.16  ||  2.11  ||  16.979  || '7959170' ||
 * 16 ||  83.36  ||  -44.51  ||  1.15  ||  0  ||  5.99  ||  0  ||  0.3  ||  0.07  ||  16.898  || '7842136' ||
 * 17 ||  82.74  ||  -44.57  ||  0.9  ||  0  ||  5.96  ||  0  ||  0.16  ||  0.01  ||  14.492  || '6431688' ||
 * 18 ||  82.62  ||  -47.77  ||  0  ||  0  ||  5.99  ||  0  ||  0.57  ||  0.04  ||  34.819  || '6565654' ||
 * 19 ||  81.55  ||  -56.82  ||  2.99  ||  0  ||  2.97  ||  0  ||  2.19  ||  2.22  ||  40.33  || '5739880' ||
 * 20 ||  81.39  ||  -43.15  ||  3.51  ||  0  ||  4.75  ||  0  ||  1.44  ||  2.08  ||  19.797  || '7825083' ||
 * 21 ||  81.34  ||  -63.98  ||  0.34  ||  0  ||  2.83  ||  0  ||  0.37  ||  0.09  ||  55.977  || '7759524' ||
 * 22 ||  81.31  ||  -35.27  ||  0.83  ||  0  ||  7.28  ||  0  ||  0.08  ||  0.01  ||  7.771  || '9040855' ||
 * 23 ||  80.96  ||  -51.71  ||  2  ||  0  ||  3.94  ||  0  ||  0.23  ||  0.07  ||  30.387  || '7724431' ||
 * 24 ||  80.51  ||  -68.93  ||  0.93  ||  0  ||  1.96  ||  0  ||  1.5  ||  0.05  ||  65.244  || '7938694' ||
 * 25 ||  79.53  ||  -37.97  ||  0  ||  0  ||  6.99  ||  0  ||  0.2  ||  0.02  ||  11.135  || '5135469' ||
 * 26 ||  79.32  ||  -48.79  ||  3.09  ||  0  ||  3.58  ||  0  ||  1.17  ||  2.11  ||  17.292  || '7964554' ||
 * 27 ||  79.31  ||  -37.43  ||  0.94  ||  0  ||  6.88  ||  1.6  ||  0.32  ||  0  ||  39.554  || '7814264' ||
 * 28 ||  79.27  ||  -56.8  ||  0  ||  0  ||  3.83  ||  0  ||  1.79  ||  3.04  ||  18.414  || '7623092' ||
 * 29 ||  79.14  ||  -50.69  ||  1.79  ||  0  ||  3.99  ||  0  ||  0.44  ||  0.05  ||  14.154  || '7589277' ||
 * 30 ||  79.04  ||  -35.68  ||  0.94  ||  0  ||  6.99  ||  0  ||  0.71  ||  0.01  ||  24.478  || '6433056' ||

__**Week of 10/20/13 - 10/26/13**__





Analysis: The activity assays don't indicate a distinct correlation between velocity and concentration such that the Km value can be determined. most of the results indicate sporadic values that are not suggestive of the trend expected. The results should indicate an increase in velocity as Nitrocefin concentration increases. initially,k we thought that the difficulty in determining the Km value came from the linear region of the Absorbance vs Time graph being too short and thus making it difficult to determine an accurate velocity. Thus, we reduced the Nitrocefin concentrations and tried the assay over a larger range of concentrations in our 2nd attempt at the activity assay. We also conducted the experiment with n=3 trials during the experiment in order to show reproducibility. However, the results of the experiment still didn't indicate the data indicative of a curve that could denote km. In order to try to increase the length of the linear region of the Absorbance vs Time curve even further such that it would be easier to determine the velocity per concentration of Nitrocefin, we reduced the enzyme concentration by 1/2, from 2ug to 1ug per 500uL of total reaction mix in the cuvette. The results still indicated poor Km relation and poor velocity vs concentration correlation.

__**Week of 10/13/13 - 10/19/13**__ Nice work Brendan. Good captions and analysis. Thank you. -Max 10/21/13 // 10/18/13: Enzyme Assay //





Analysis: Reduced the concentration of working dilution protein and increased the volume. Also halved the concentration of protein in order to increase the length of the linear region of the curve. However, the data still shows sporadic data that indicates no distinct trend. More assays to come!

// 10/14/13: Enzyme Assay //



Analysis: Better results, but using GraphPad Prism (results to be shown next week due to lack of access), the Km values were determined to be nonsense (>118). It is suspected that inconsistencies in stirring the protein into the nitrocefin could be the cause of the problem. In order to reduce this issue, another activity assay will be conducted that uses a larger volume of a more dilute working dilution of NDM-1 protein, so it hopefully disperses into the rest of the solution more evenly.

__**Week of 10/6/13-10/12/13:**__

//10/7/13: Enzyme assay// __Thoughts on Enzyme Assay:__ Experiment needs to be redone. The values received from the 20uM and 38.5uM nitrocefin assays are as expected, namely, that the slope or velocity of the reaction increased for the greater concentration of substrate. However, this trend seemingly discontinued at the higher concentrations of 5.56uM and 100uM nitrocefin.

//10/7/13 - Control Ligand Docking in ICM Conducted// controls outranked some of the most notable positive controls such as ampicillin.
 * NAME || Score || Natom || Nflex || Hbond || Hphob || VwInt || Eintl || Dsolv || SolEl || mfScore || POS || RecConf ||
 * 5742832aztreonampos4 || 33.972006 || 45 || 7 || -4.279057 || -3.652 || -24.2 || 0.325956 || 18.418354 || 80.434619 || -80.5391 || 14 || 1 ||
 * 2244asprin || 30.948482 || 21 || 1 || -2.977612 || -3.325 || -11.9 || 0 || 9.203528 || 68.359647 || -75.1042 || 10 || 1 ||
 * 65894faropenempos7 || 27.766007 || 34 || 4 || -1.899556 || -4.221 || -18.8 || 0 || 12.940278 || 63.800214 || -103.736 || 4 || 1 ||
 * 5280980clavulanicpos5 || 20.04634 || 23 || 4 || -1.220113 || -3.187 || -14.1 || 0 || 10.577421 || 44.021473 || -65.5747 || 15 || 1 ||
 * 5904penicillingpos10 || 14.989494 || 41 || 5 || -8.425242 || -4.076 || -22.9 || 0.082477 || 14.850178 || 72.325838 || -113.689 || 1 || 1 ||
 * 5762077neg1 || 14.030338 || 40 || 2 || -4.134254 || -5.283 || -29.9 || 1.616009 || 12.804248 || 68.938438 || -74.9564 || 8 || 1 ||
 * 1033244neg3 || 11.40441 || 52 || 4 || -1.032907 || -7.052 || -31.4 || 0 || 7.872336 || 63.349512 || -61.8169 || 6 || 1 ||
 * 17399864neg4 || 10.794869 || 37 || 2 || 0 || -5.827 || -22.7 || 1.968481 || 8.710066 || 41.929861 || -60.2192 || 9 || 1 ||
 * 20824carbenicillin || 10.145432 || 44 || 7 || -4.383559 || -4.716 || -22 || 1.036014 || 16.204445 || 43.900916 || -89.0735 || 12 || 1 ||
 * 71339biapenempos6 || 9.824641 || 42 || 4 || -1.211114 || -4.986 || -18.5 || 0 || 13.946948 || 33.116053 || -105.377 || 5 || 1 ||
 * 33255Cefazolinpos8 || 9.325897 || 43 || 6 || -9.472784 || -5.12 || -31 || 0.518731 || 24.383932 || 68.627518 || -126.68 || 3 || 1 ||
 * 6436055cefcapenepos3 || 9.280157 || 49 || 5 || -7.642363 || -4.923 || -33.8 || 0 || 21.734659 || 70.289256 || -114.487 || 13 || 1 ||
 * 6453491ampicillinoicacidpos1 || 7.550721 || 46 || 9 || -8.235407 || -4.123 || -21.8 || 1.027698 || 19.700354 || 48.151559 || -72.9888 || 11 || 1 ||
 * 6249ampicillinpos9 || 4.978397 || 43 || 6 || -8.404371 || -4.396 || -21.9 || 0.99322 || 15.235351 || 53.13562 || -80.6112 || 2 || 1 ||
 * 3129153neg2 || 4.7057 || 45 || 3 || -3.278469 || -5.908 || -30.8 || 0.593811 || 13.714974 || 53.132364 || -112.503 || 7 || 1 ||
 * Figure 1:** Control ligand docking to NDM-1 betalactamase conducted in ICM. Results are strange because some of the negative

__Analysis:__ Need to take another look at the reasons control docking of these ligands occurs strangely. The top few ligands make sense in the sense that aztreonam and clasvulanic acid are the only two drugs known to possibly inhibit NDM-1 betalactamse. However, it is not yet clear why these negative controls outranked ligands such as ampicillin, because NDM-1 betalactamse cleaves the beta-lactam ring of ampicillin.

__**Week of 9/30/13-10/5/13:**__
 * Good captions but try and add an analysis. Thank you. -Max 10/07/2013

//10/5/13:// ChemBridge docking to 4EY2 begun. scored very highly. The Autoscale was set to 2.
 * # || Score || S(PLP) || S(hbond) || S(cho) || S(metal) || DE(clash) || DE(tors) || intcor || time || Ligand name ||
 * 1 || 92.78 || -57.09 || 3.26 || 1 || 3.96 || 0 || 1.29 || 1.59 || 26.476 || '33255Cefazolinpos8' ||
 * 2 || 91.96 || -74.85 || 0 || 0 || 2.98 || 0 || 0.41 || 0.04 || 26.104 || '3129153neg2' ||
 * 3 || 91.8 || -74.59 || 0 || 0 || 2.99 || 0 || 0.41 || 0.03 || 46.328 || '3129153neg2' ||
 * 4 || 91.26 || -55.82 || 3.18 || 1 || 3.94 || 0 || 1.22 || 1.65 || 26.749 || '33255Cefazolinpos8' ||
 * 5 || 90.81 || -71.61 || 0 || 0.94 || 3 || 0 || 0.84 || 0.06 || 27.16 || '3129153neg2' ||
 * 6 || 90.41 || -70.42 || 0 || 1 || 2.97 || 0 || 0.45 || 0.04 || 21.168 || '3129153neg2' ||
 * 7 || 89.98 || -70.67 || 0 || 0.54 || 3 || 0 || 0.16 || 0.04 || 19.195 || '3129153neg2' ||
 * 8 || 89.78 || -51.55 || 1.36 || 0 || 5.85 || 0 || 0.64 || 0.3 || 27.678 || '6453491ampicillinoicacidpos1' ||
 * 9 || 89.08 || -72.89 || 0 || 0 || 2.93 || 0 || 0.73 || 0.08 || 18.626 || '3129153neg2' ||
 * 10 || 88.6 || -52.71 || 1 || 0 || 5.97 || 0 || 1.66 || 0.38 || 28.928 || '6453491ampicillinoicacidpos1' ||
 * 11 || 86.86 || -46.4 || 0.52 || 0 || 6.95 || 0 || 1.65 || 0.52 || 13.904 || '65894faropenempos7' ||
 * 12 || 86.73 || -46.1 || 0.63 || 0 || 6.91 || 0 || 1.61 || 0.48 || 13.571 || '65894faropenempos7' ||
 * 13 || 86.62 || -46.13 || 0.8 || 0 || 6.84 || 0 || 1.72 || 0.5 || 14.763 || '65894faropenempos7' ||
 * 14 || 83.74 || -44.1 || 1.57 || 0 || 5.96 || 0 || 0.66 || 0.49 || 61.672 || '20824carbenicillin' ||
 * 15 || 83.36 || -46.09 || 0.54 || 0 || 5.99 || 0 || 0.33 || 0.38 || 62.947 || '6249ampicillinpos9' ||
 * 16 || 80.1 || -46.93 || 3.8 || 1 || 3.83 || 0 || 3.31 || 2.44 || 96.139 || '6436055cefcapenepos3' ||
 * 17 || 80.03 || -47.92 || 3.12 || 1 || 3.96 || 0 || 3.39 || 2.74 || 99.119 || '6436055cefcapenepos3' ||
 * 18 || 79.4 || -79.92 || 0 || 0 || 0 || 0.25 || 2.22 || 4.15 || 46.051 || '1033244neg3' ||
 * 19 || 79.2 || -49.01 || 2 || 1 || 3.97 || 0 || 1.43 || 0.22 || 19.093 || '71339biapenempos6' ||
 * 20 || 72.12 || -30.39 || 0.86 || 0 || 6.97 || 0 || 1.34 || 0.01 || 11.773 || '2244asprin' ||
 * 21 || 71.95 || -33.5 || 0.86 || 0 || 5.99 || 0 || 0.09 || 0.09 || 10.979 || '5280980clavulanicpos5' ||
 * 22 || 70.09 || -46.1 || 4.42 || 0 || 1.97 || 0 || 2.45 || 3.81 || 64.004 || '5742832aztreonampos4' ||
 * 23 || 69.94 || -64.97 || 0 || 0 || 0.97 || 0 || 0.42 || 0.01 || 13.793 || '5762077neg1' ||
 * 24 || 68.43 || -71.65 || 0 || 0 || 0 || 2.89 || 0.36 || 0.38 || 31.331 || '17399864neg4' ||
 * 25 || 67.4 || -51.31 || 1.84 || 0 || 1.96 || 0 || 0.72 || 0.25 || 19.105 || '5904penicillingpos10' ||
 * 26 || 61.51 || -58.98 || 1 || 0 || 0 || 0 || 0.24 || 0 || 9.694 || '17399864neg4' ||
 * Figure 1:** Control Docking results from GOLD. 26 ligand-prepped results shown. One of the negative controls

//10/4/13//: Control Docking completed. Begin ligand prep on the ChemBridge Library. //10/1/13:// Ligand Control prep completed for virtual //9/31/13//: Protein Prep completed for Virtual

__**Week of 9/23/13-9/29/13:**__ // 9/26/13: // FPLC


 * Figure 1:** Nanodrop of Elution 1 of NDM-1 protein after Ni-NTA purification, prior to FPLC protocol.


 * Figure 2:** FPLC protocol graphical analysis. Tubes 39-48 taken for reconcentration.




 * Figure 3:** Nanodrop results of NDM-1 Elution 1 after FPLC purification.


 * Week 3 & 4 9-Sep - 22-Sep**
 * Brendan - good work. Move forward to enzyme assays. Dr. B 100113**
 * PS - interesting grid on your master plate...?**

__**Week of 9/16/13-9/22/13:**__

//September 19, 2013//: Continue washing off excess dye. Need to FPLC due to the presence of other bands. **Figure 1:** Gel of protein NDM-Beta-lactamase after purification. Lanes from left to right: Ladder, Flow through, Wash, Elution 1, Elution 2, Soluble Fraction. The ColorPlus protein ladder is shown to the below with the different MWs shown beside each band.

Gel looks pretty successful. the band is darkest for the Elution 1 sample and is considerably lighter for the Elution 2 sample. There are still significant bands at other MW, so FPLC will be performed for further purification starting next week.

The protein target has a molecular weight of 28kDa. This seems to be where the ladder indicates the darkest band is showing up in the Elution samples.

//September 18,// //2013:// Ran gel of protein. Dyed and now washing off excess dye. Tomorrow = imaging. //September 16, 2013:// Purification and storage. Next up: //characterization.//

__**Week of 9/9/13-9/15/13**__ //September 13, 2013:// Lysing of cells occurred. After lysing, stored in fridge after spin down and initial filtration to await next week's purification

//September 12, 2013:// Centrifuge down of the cells at 4 degrees C and 6300 rpm. Glycerol stocks made and placed in cryovials in the -80C fridge. Tube 1: 3.15g Tube 2: 1.98g

//September 11, 2013//: Relatively few cultures of bacteria seen, this is expected. Started culturing after OD600 of ~0.5 was reached. Kan was added late, and OD600 was closer to 0.8 than 0.5 when placed in a room temperature shaker for overnight. Glycerol Stocks protocol Day 1 started. Transfer to cryovials tomorrow.



plates. 50uL of culture added to plate.
 * Figure 1:** Plated B21(DE3) E.Coli transformed with p27bNDM(35a) on LB+Agar+Sucrose



plates. 10uL of culture added to plate.
 * Figure 2:** Plated B21(DE3) E.Coli transformed with p27bNDM(35a) on LB+Agar+Sucrose

//September 10, 2013//: Only a few colonies remaining. Perhaps 30uL/mg as indicated by Fast lab protocol should be taken into advisement. However, this could also have been a product of the timing after heat shocking. Made two culture flasks, one with 30uL/mL Kan, and the other with 50uL/mL Kan in case the 50uL/mL Kan culture doesn't turn out turbid. // September 9, 2013 // : Transformation on to Kan plates. Made fresh LB for the next day's culturing. Note: Didn't allow for full 2min after heat-shocking before adding SOC media.

__**Week of 9/2/13-9/8/13**__ //Good - Dr. B 090913//

//September 6,// 2013: Target Group Meeting. __To-Do List:__ //1. Wet Lab// -Transformation+Expression [ []] >>
 * 9/9 - Day 1 : Transformation+Make LB [ []]
 * 9/10 - Day 2 : Take colonies and make cultures with 30ug/mL Kan.
 * 9/11 - Day 3 <3.5hrs>: OD600 testing [37C]-->Induction-->18-20hrs @ 25C growth.
 * Make glycerol stocks prior to other stuff : [[|https://docs.google.com/file/d/0B_Gl3lMNitryhDsoc0FHRXhRRnJDSkk/edit?usp=sharing]]
 * 9/12 - Day 4 : Collect cells: Spin for 20 min at 6000 g. [Can store for days in -20C, or for hours in 4C]
 * 9/13 - Day 5 <1.5hrs>: Reserve and precool centrifuge. Use Option A Lysis protocol-->Sonicate spin @ 16k rpm, 30min

- 9/16 - Purification <2hrs-2.5hrs> [[]] [store in 4C] - 9/18 - Characterization [[]] > - Preliminary assay - Nitrocefin at varying concentrations
 * 9/13 - Make SDS-PAGE Gel soln 1-2 days early <1.5hr> : [[]]

//2. Virtual -// with the aid of the gracious Madeline :D -GOLD screen preliminary library:
 * LabVS4 NDM-1 Redo: [h[|ttps://docs.google.com/file/d/0B_Gl3lMyhDsoYnVCcWhOVEJoQkE/edit?usp=sharing]]
 * Ligand Prep: [[]]

-ICM/VINA Docking, ICM Screen certain attributes and compare results to GOLD ligand docking vs attribute-filtered library -GraphPad Prism - enzyme kinetics determination

//September 4, 2013:// Results from DNA-Sequencing received and analyzed. Blastn searches run for both of these sequences.

T7:  code NNNNNNNNNNNNCNTCTAGANTAATTTTNTTTAACTTTAAGAAGGAGATATACATATGAAATACCTGCTGCCGACCGCTG CTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCATGGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGC GATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGC TTCCAACGGTTTGATCGTCAGGGATGGCGGCCGCGTGCTGGTGGTCGATACCGCCTGGACCGATGACCAGACCGCCCAGA TCCTCAACTGGATCAAGCAGGAGATCAACCTGCCGGTCGCGCTGGCGGTGGTGACTCACGCGCATCAGGACAAGATGGGC GGTATGGACGCGCTGCATGCGGCGGGGATTGCGACTTATGCCAATGCGTTGTCGAACCAGCTTGCCCCGCAAGAGGGGAT GGTTGCGGCGCAACACAGCCTGACTTTCGCCGCCAATGGCTGGGTCGAACCAGCAACCGCGCCCAACTTTGGCCCGCTCA AGGTATTTTACCCCGGCCCCGGCCACACCAGTGACAATATCACCGTTGGGATCGACGGCACCGACATCGCTTTTGGTGGC TGCCTGATCAAGGACAGCAAGGCCAAGTCGCTCGGCAATCTCGGTGATGCCGACACTGAGCACTACGCCGCGTCAGCGCG CGCGTTTGGTGCGGCGTTCCCCAAGGCCAGCATGATCGTGATGAGCCATTCCGCCCCCGATAGCCGCGCCGCAATCACTC ATACGGCCCGCATGGCCGACAAGCTGCGCGCTAGCCAGCCAGAACTCGCCCCGGAAGACCCCGAGGATGTCGAGCACCAC CACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGANTTGGCTGCTGCCACCGCTGANCAATAACT AGCATAACCCCTTGGGGNNCTAAACGGGTCTTGNNGGGTTTTTGCTGAAGGNNNNTATATCCGNATTGGCGAATGGGNNG NNNCNGNAGNNNGCATTNAGNNNGNNNNNNGNNNNCNCNCANCNNNACNCTANNCTTNCNGCNNNANNNCCNNNNNTTCN NTTTCNTNNNNNTNNNNNNCNNNNNTNNNNNNTTTNCNNNANNNNNAANNGGGGNNCNCNNNNNGNNNNNNTNNNNNNNN GNNNNCNNNNNNNNANANNTNNNNNNNNGNNNNNNNNNNNNNNNNNNNNNN code **Figure 1:** DNA-Sequencing results from CORE after Midi-prep of pET-27b(+) NDM-1(d35) plasmid. Shown with the  T7 in-house primer.  T7Term  code NNNNNNNNNNNNNNNTTCGGGCTTTGTTAGCAGCCGGATCTCAGTGGTGGTGGTGGTGGTGCTCGACATCCTCGGGGTCT TCCGGGGCGAGTTCTGGCTGGCTAGCGCGCAGCTTGTCGGCCATGCGGGCCGTATGAGTGATTGCGGCGCGGCTATCGGG GGCGGAATGGCTCATCACGATCATGCTGGCCTTGGGGAACGCCGCACCAAACGCGCGCGCTGACGCGGCGTAGTGCTCAG TGTCGGCATCACCGAGATTGCCGAGCGACTTGGCCTTGCTGTCCTTGATCAGGCAGCCACCAAAAGCGATGTCGGTGCCG TCGATCCCAACGGTGATATTGTCACTGGTGTGGCCGGGGCCGGGGTAAAATACCTTGAGCGGGCCAAAGTTGGGCGCGGT TGCTGGTTCGACCCAGCCATTGGCGGCGAAAGTCAGGCTGTGTTGCGCCGCAACCATCCCCTCTTGCGGGGCAAGCTGGT TCGACAACGCATTGGCATAAGTCGCAATCCCCGCCGCATGCAGCGCGTCCATACCGCCCATCTTGTCCTGATGCGCGTGA GTCACCACCGCCAGCGCGACCGGCAGGTTGATCTCCTGCTTGATCCAGTTGAGGATCTGGGCGGTCTGGTCATCGGTCCA GGCGGTATCGACCACCAGCACGCGGCCGCCATCCCTGACGATCAAACCGTTGGAAGCGACTGCCCCGAAACCCGGCATGT CGAGATAGGAAGTGTGCTGCCAGACATTCGGTGCGAGCTGGCGGAAAACCAGATCGCCAAACCGTTGGTCGCCAGTTTCC ATTTGCTGGCCCATGGCCATCGCCGGCTGGGCAGCGAGGAGCAGCAGANCAGCAGCAGCGGTCGGCAGCAGGTATTTCAT ATGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAGAGGGGAATTGNTATCCGCTCACAATTCCCCTATAGTGAG TCGTATTAATTTCGCGGGATCGAGATCTCGATCCTCTACNCCGNANGCATCGNGGNNGGCATCACCGGNNCCNNNGNTGC NGNTGCTGGCGNCTATATCNNNANNTCNNCNAATGGGGNNNNTNGGGCNCGCCNCTTNGNCTCNTGNANNNNTGTTCNGN NNNNGGGNNNGGNNGNNGNCCCNNNNNNNNNNTNNNNGNNNCNTCNCNTNNNNNNNNNCNNNNNNNCGNNGGNGGNNNNN ANNGNNNNCNNCNNNNNGNNNNNNTTNNANNNANNANTNNNNNNNNNNNANNNNNNNNNNNNNNNNNNNNNNNNNGN code **Figure 2:** DNA-Sequencing results from CORE after Midi-prep of pET-27b(+) NDM-1(d35) plasmid. Shown with the  T7term in-house primer.   <span style="background-color: #ffffff; color: #222222; display: block; font-family: arial,sans-serif; font-size: 12.7273px;">

confirms the presence of the <span style="background-color: #ffffff; color: #222222; font-family: arial,sans-serif; line-height: 1.5;">pET-27b(+) NDM-1(d35) plasmid.
 * Figure 3:** Blastn search results of the DNA sequence received from the CORE. Blastn search (of T7Term primer+plasmid)



confirms the presence of the <span style="background-color: #ffffff; color: #222222; font-family: arial,sans-serif;">pET-27b(+) NDM-1(d35) plasmid.
 * Figure 4:** Blastn search results of the DNA sequence received from the CORE. Blastn search (of T7 primer+plasmid)

//September 2, 2013//: Midi prep of DH5a transformed with pET-27b(+) NDM-1(d35). And nanodrop of results. Plasmids sent for sequencing using the T7term and T7 in-house primers provided by the CORE.

Figure 1: Nanodrop of p276-NDM(35) after Midi-prep. 260/230 <span style="font-family: Calibri,sans-serif; font-size: 11pt;">values are too low, and the graph indicates a negative 230 value.

Figure 2: Nanodrop of p276-NDM(35) after Midi-prep. 260/230 values are too high, and the graph indicates a negative 230 value.



Figure 3: Nanodrop of p276-NDM(35) after Midi-prep. 260/230 values are now as Expected. Used new blanking TE Buffer and new nanopure water for blanking and calibrating.



Figure 4: Nanodrop of p276-NDM(35) after Midi-prep. 260/230 values are now as expected. Used new blanking TE Buffer and new nanopure water for blanking and calibrating.

__**Week of 8/28/13 - 9/1/13:**__

//August 31, 2013//: Spin down and fridge colonies after 20hrs. //August 30, 2013:// Took two colonies and incubated overnight after autoclaving LB media. //August 29, 2013:// Transformed DH5a with pET27b(+) NDM-1(d35) on Kan plates, for later Midi-prep. Running low on plasmid.


 * __Week 3:__**

Protein expression completed.




 * Figure 1:** Blast results of GFP plasmid sent for sequencing.



Lane 3 and 8 = 5ng GFP, Lane 4 and 9 = 25ng GFP. Lane 5 and 10 = 0ng. Lanes 1-5 with PCR with M13 primers, at M13 primer optimal temperatures, and Lanes 6-10 with PCR with VDS1 and VDS2 primers, at M13 primer optimal temperatures.
 * Figure 2:** Gel electrophoresis of GFP DNA post-PCR. Lane 1 and 6 = Ladder, Lane 2 and 7 = 1ng GFP,


 * __Week 2:__**



was sent to sequencing to confirm Midi-prep results.
 * Figure 1:** Nanodrop measurement of pNIC-Bsa4 after PCR. Concentration of 301.3 ng/uL exhibited. A sample of this plasmid solution



was sent to sequencing to confirm Midi-Prep results.
 * Figure 2:** Nanodrop measurement of pNIC-Bsa4 after PCR. Concentration of 283.0 ng/uL exhibited. A sample of this plasmid solution

Lane 6 = Ladder, Lane 7 = 1ng pgbr22, Lane 8 = 5ng pgbr22, Lane 9 = 25ng pgbr22. Lane 10 = 0ng Lanes 1-5=Nicolet’s.
 * Figure 3:** Brendan (Right) pgbr22 post-PCR. About 1kb.

__ **Week 1** __ Figure 1: E. coli cells transformed with 1 ng pNIC-Bsa4 plasmid grown on LB+Amp Agar plates. Approximately 1648 colonies/ng plasmid.

Figure 2: E. coli cells transformed with 5 ng pNIC-Bsa4 plasmid grown on LB+Amp Agar plates. Approximately 3072 colonies/ng plasmid.

Figure 3: E. coli cells transformed with 25 ng pNIC-Bsa4 plasmid grown on LB+Amp Agar plates. Approximately 689 colonies/25ng, = 27.6coloines/ng plasmid.