Dax+F.+G.

Week 15 Enzyme assay and inhibition assays were done this week. Virtual was attempted, but due to fault in preparation of files, was not able to run on the processors and yield any results.









Week 14 On this week, expression, purification, FPLC, characterization, and storage were completed.

















Week 13 On this week, mini-prep and submitting to DNA sequencing were performed. The DNA sequencing results came back blank or with very, very few codons. My partner's data came back similarly. Thus it was decided that a surrogate target, YopH, would be pursued. Expression was started with the surrogate.













Good. Include pics of your nanodrops after mini prep -UM Week 12 After the second attempt at cloning failed, the first master plate was returned to, and more attempts to grow liquid cultures were made. First, a fresh batch of LB and agar plates were made from scratch, and immediately used, with the contents double checked to ensure that the plates and LB being used were not causing the cultures lack of growth. Then, fresh cultures from the original plates, labeled as "B" were chosen and made a master plate with, and the cultures from the first master plate were re-plated and labeled "A". When all plated cultures grew, but the liquid cultures did not, the liquid cultures were grown again at room temperature. The liquid cultures that grew at room temperature were successful, and mini-prep was performed.





All 12 "A" colonies, and 8 "B" colonies were used to make fresh master plates, pictured above, and were put into liquid broth in order to be grown. All 20 colonies failed in the 37 degree incubator during the first round. Then eight colonies, (1A, 4A, 7A, 10A, 8A, 11A, 1B and 2B) were selected from the new master plates and grown at room temperature for 24 hours, and did grow successfully.





Week 11 For this week, cloning was continually attempted with various components changed in order to attempt to grow a viable set of cultures.

The second attempt at cloning did not produce an original plate as the cultures did not grow.





Notes: The liquid cultures are not pictured because they did not show any growth. It was determined after that step of cloning was completed the first time that too much Kanamycin had been added, and that had probably killed off the cultures. Using the master plate, twelve new LB tubs were incubated with the cultures, however none of these grew either. This step was repeated a third time, this time warming the LB, but bacterial growth failed once again. A second round of cloning was started in the meantime to try to prevent anymore lost time.



Where is Week 10? Missing plate pic. -UM Week 9 and 10 For weeks 9 and 10, the first round of cloning was performed. To minimize chance for error, the protocol was followed on back to back days until the end of the master plate protocol. However, it was found at the very end of this step that the tubes from the master plate protocol had received too much Kanamycin and will have to be redone.

There is not yet a picture of the master plate, but it should be noted that while the colonies in the tubes did not grow, there was growth in all twelve sectors of the master plate.





Week 8 For week 8, midiprep and the transformation of pNIC were done. The first transformation was completed at the end of week 7, and then the first midiprep, second transformation of pNIC and second midiprep were all completed within week 8.

The second attempt at midiprep proved even worse for yield, and was not fit to move on into cloning. Because of time, at this point midiprep was abandoned and another sample from a midiprep performed by Will E. was used for cloning in week 9.





With the low yield produced by midiprep, it was decided that transformation and midiprep should be repeated for a better yield.







Very good! Have you tried Gel extraction to get rid of that contam. from pcr squared? -UM Week 7 Work on PCR was continued throughout this week, and transformation of PNIC was started.

The fourth round of squared was cleaned up and nanodropped, and showed a high yield.

The fourth round of squared was finally successful.

The third attempt at squared failed because of a gel error.

The first two rounds of squared were not considered successful because the bands were not bright enough.

The fourth attempt at secondary was finally successful.

Good images and captions. Very good analysis! Now just cut back on the human errors and hopefully things will work better! - Michael T Week 5 & 6

PCR for the Target Vibrio Cholerae was run throughout the two weeks. During this time period, dilutions, LB, and LB agar plates were also made for future use.

Third attempt at Secondary: Secondary was attempted again with the same primary sample. For this round of secondary, the temperature during the annealing step was raised by two degrees, as suggested by William (also working on the target) who used those qualifications to get his secondary to work. The times were also adjusted to pick the middles times for each of the steps, to shorten the steps in order to try to get secondary to work. Ultimately, the times were probably shortened too much, because nothing showed up at all on this gel run for this attempt at PCR. It is also suspected that there was a problem with the gel because of the distortion as seen in the image. It should be noted that while the lane that was supposed to contain secondary did not show up on the image, when looking at the gel directly, a blue marker could be seen in the second lane.



Second attempt at Primary and Secondary PCR: Primary and secondary were both run before putting on a gel a second time, this time, it was noted that the wrong times had been used in the first attempt, and the correct times were used. Primary PCR left a smear and worked, but secondary smeared as well, indicating it did not work.



First attempt at Primary and Secondary PCR: Primary and secondary were both run before putting on a gel. Due to pipette error, all of each of the samples was taken out to mix with blue dye for a run on the gel. Once the gel was run, it was discovered that someone had mixed in ladder to the blue dye, and so all of the primary and secondary samples were contaminated, and had to be redone.

Week 3 & 4 Dax - crop your image in the imaging software, include a ladder image. Missing a Primary PCR run. Otherwise - looks good. - Dr. B 092713

Week 4 Second PCR of pGbr

Note that sample D was accidentally placed in lane 2, and sample A was placed in lane 5.

Analysis: This is a successful PCR of pGbr 22, showing nothing in lane 2, where sample D, without DNA is located, and gaining brightness going from sample A to C.

Week 3 First PCR of pGbr

Analysis: The first PCR pf pGbr22 failed, with sample D being the only lane that showed, and sample D supposedly being the sample not containing DNA. It is suspected that cross contamination is the cause.

RE Digest of pGbr 22 Analysis: Compared with the ladder, we can see only a smear with the uncut plasmid, and different markers for where each of the following samples cut the DNA sequence, with the sections being compiled in the last lane that contains both EcoRI and PvuII.

Primer Tail Design:









Phosphoserine Phosphotase Vibrio Cholerae Primer Tale Design

Sequence to be inserted into PNIC, with the start and end highlighted in green, and the tails highlighted in yellow.

TACTTCCAATCC ATGGACGCGCTGACCA CCCTCCCGATCAAAAAGCACACCGCGCTGCTGAACCGTTTC CCGGAAACCCGCTTCGTTACCCAACTGGCGAAAAAGCGTGCGTCTTGGATCGTTTTCGGTCAC TACCTCACTCCAGCACAGTTTGAAGATATGGATTTTTTCACCAATCGTTTCAATGCGATC CTGGACATGTGGAAAGTTGGCCGTTACGAAGTTGCGCTGATGGACGGTGAACTGACCTCT GAACACGAAACCATCCTGAAAGCGCTGGAACTCGACTACGCTCGCATCCAGGACGTTCCA GACCTCACCAAACCGGGCCTGATCGTTCTCGACATGGACTCTACCGCTATCCAGATCGAA TGCATCGACGAAATTGCGAAGCTGGCGGGTGTTGGCGAGGAAGTGGCCGAAGTTACGGAA CGTGCGATGCAGGGCGAGCTGGACTTCGAACAGTCTCTGCGTCTGCGTGTTTCTAAACTC AAAGACGCCCCTGAACAGATCCTGAGCCAGGTTCGTGAAACGCTGCCGCTCATGCCTGAA CTGCCGGAACTGGTTGCGACCCTGCACGCGTTCGGTTGGAAGGTAGCAATCGCGTCTGGT GGTTTCACCTACTTTTCTGACTACCTGAAGGAACAACTCAGCCTCGATTACGCGCAGTCT AACACCCTGGAAATTGTTTCTGGTAAACTGACTGGTCAAGTTCTGGGTGAAGTTGTGTCT GCTCAGACCAAAGCGGACATCCTGCTGACCCTGGCGCAACAGTACGACGTTGAAATCCAC AACACCGTTGCGGTGGGTGACGGTGCGAACGACCTGGTTATGATGGCGGCTGCGGGCCTC GGTGTAGCGTACCATGCGAAACCGAAGGTTGAGGCGAAGGCGCAGACCGCAGTTCGTTTC GCTGGTCTCGGTGGTGTCGTTTGCATCCTGTCTGCGGCGCTCGTTGCGCAGCAAAAA CTC TCTTGGAAATCTAAACCGTAA CAGTAAAGGTGGATA

Forward Primer: 30 bp GC content: 56.7%

Reverse Primer: 33 bp GC content: 36.4%


 * mM of Mg2+ in Tm °C || Forward || Reverse ||
 * 0 || 66.8 || 59.3 ||
 * 1.5 || 73.1 || 67.2 ||
 * 2 || 73.6 || 67.7 ||
 * 4 || 74.5 || 68.8 ||
 * 6 || <span style="background: none repeat scroll 0% 0% white; color: #000000; line-height: normal; margin-bottom: 0in;">74.9 || <span style="background: none repeat scroll 0% 0% white; color: #000000; line-height: normal; margin-bottom: 0in;">69.3 ||

Note that base pairs were added and removed from both ends until the 2mM of Mg2+ had temperatures of 72.4° and 69.8° Celsius for the forward and reverse primers respectively.

Primers to Order:

__ Upstream: __

TACTTCCAATCCATGGACGCGCTGACCA

__ Downstream (reverse complemented): __

TATCCACCTTTACTGTTACGGTTTAGATTTCCAAGAGAG

*Note that this insert contains code that codes for a cut by BsaI.

__ Full pNIC-Bsa4 sequence with insert, start and end highlighted in green, and the 6-His tag highlighted in yellow. __

<span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATG CACCATCATCATCATCAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TCTTCTGGTGTAGATCTGGGTACCGAGAACCTG <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">TACTTCCAATCC ATGGACGCGCTGACCA CCCTCCCGATCAAAAAGCACACCGCGCTGCTGAACCGTT <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">TCCCGGAAACCCGCTTCGTTACCCAACTGGCGAAAAAGCGTGCGTCTTGGATCGTTTTCGGTCACTACCTCACTCCAGCACAGTTTGAAGATATGGA <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">TTTTTTCACCAATCGTTTCAATGCGATCCTGGACATGTGGAAAGTTGGCCGTTACGAAGTTGCGCTGATGGACGGTGAACTGACCTCTGAACACGAA <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">ACCATCCTGAAAGCGCTGGAACTCGACTACGCTCGCATCCAGGACGTTCCAGACCTCACCAAACCGGGCCTGATCGTTCTCGACATGGACTCTACCG <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">CTATCCAGATCGAATGCATCGACGAAATTGCGAAGCTGGCGGGTGTTGGCGAGGAAGTGGCCGAAGTTACGGAACGTGCGATGCAGGGCGAGCTGGA <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">CTTCGAACAGTCTCTGCGTCTGCGTGTTTCTAAACTCAAAGACGCCCCTGAACAGATCCTGAGCCAGGTTCGTGAAACGCTGCCGCTCATGCCTGAA <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">CTGCCGGAACTGGTTGCGACCCTGCACGCGTTCGGTTGGAAGGTAGCAATCGCGTCTGGTGGTTTCACCTACTTTTCTGACTACCTGAAGGAACAAC <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">TCAGCCTCGATTACGCGCAGTCTAACACCCTGGAAATTGTTTCTGGTAAACTGACTGGTCAAGTTCTGGGTGAAGTTGTGTCTGCTCAGACCAAAGC <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">GGACATCCTGCTGACCCTGGCGCAACAGTACGACGTTGAAATCCACAACACCGTTGCGGTGGGTGACGGTGCGAACGACCTGGTTATGATGGCGGCT <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">GCGGGCCTCGGTGTAGCGTACCATGCGAAACCGAAGGTTGAGGCGAAGGCGCAGACCGCAGTTCGTTTCGCTGGTCTCGGTGGTGTCGTTTGCATCC <span style="background: none repeat scroll 0% 0% white; color: #000000; font-family: 'Courier New'; font-size: 11pt; line-height: 115%;">TGTCTGCGGCGCTCGTTGCGCAGCAAAAA CTCTCTTGGAAATCTAAACCGTAA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CAGTAAAGGTGGATACGGATCCGAATTCGAGCTCCGTCGACAAGCTTG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATTGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAATTAATTCTTAGAAAAACTCATCGAGCATCAAATGA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AACTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTTCCATAGGATGGCAAGATCCT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">ATTGCGCCTGAGCGAGACGAAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAATATT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TTCACCTGAATCAGGATATTCTTCTAATACCTGGAATGCTGTTTTCCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGATGGTCG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GAAGAGGCATAAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCAACGCTACCTTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CCCATACAATCGATAGATTGTCGCACCTGATTGCCCGACATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTAGAGCAAG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">ACGTTTCCCGTTGAATATGGCTCATAACACCCCTTGTATTACTGTTTATGTAAGCAGACAGTTTTATTGTTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">ACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATATGGTGC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">ACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGAAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGGACCAGAGAAAAATCACTCAGG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GTCAATGCCAGCGCTTCGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCAGCAGCATCCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TTCCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGTCGCAGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGATTCAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACGACAGGAGCACGATCATGCGCACCCGTGGGGCCGCCATGCCGGCGATAATGGCCTGCTTCTC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GCCGAAACGTTTGGTGGCGGGACCAGTGACGAAGGCTTGAGCGAGGGCGTGCAAGATTCCGAATACCGCAAGCGACAGGCCGATCATCGTCGCGCTCCAGCGAAAGC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GGTCCTCGCCGAAAATGACCCAGAGCGCTGCCGGCACCTGTCCTACGAGTTGCATGATAAAGAAGACAGTCATAAGTGCGGCGACGATAGTCATGCCCCGCGCCCACCG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GAAGGAGCTGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAAC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTAACGGCGGGATAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AACATGAGCTGTCTTCGGTATCGTCGTATCCCACTACCGAGATATCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGAATTTGAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGAACTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGATGCTC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACAG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CGACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAAC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">GACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCAC <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CACGCGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCA <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">TACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCG <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">AGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGAT <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%;">CCCGCGAAAT

Week 1 & 2 Dax - good job. Need your PCR results Include a one sentence analysis for each exepriment. Dr. B 090913 Analyzing DNA:







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Oligo Design:

Week 1

Nanodrop: The known concentration was listed at 205.4 ng/uL. The average concentration found was 196.7 ng/uL. Note that the measurements were taken several more times than listed to ensure the measurement was correct.