D'Ondria+P.

__**Week 9 & 10**__
__**Virtual Screening (to be continued...)**__ Figure 23 The image above is of the 10 positive and 5 negative ligand that will be used for virtual screening. __**Cut pNic-Bsa4**__ Figure 22 The image above is a 1% agarose gel loaded with 1XTAE buffer. Lane one contains 1Kb ladder, lane two contains 100bp ladder, lane three contains my 10uL PCR clean-up sample mixed with 2uL of blue dye, lane four is empty, lane five contains Brandy's sample. The 1Kb ladder reference is to the left of the gel image.
 * Good analysis and nice amount of data. Work on how to write captions for images. Also missing sample ID for nanodrop images. - BN**

__Analysis:__ The STP1 gene had 739bp. The cut sites seem to be accurate and Brandy's are in the same spot as mine which backs up that the cut sites are accurate. My sample is not as bright as Brandy's is which means that she had more of the sample than I do. Hopefully this wont mess my cloning up in the long run. The 100bp was added out of habit but it wasn't necessary. __**Cut sites**__ Figure 21 The figure above is of the NEB cut image for the restriction enzyme BsaI for pNIC-Bsa4. __**pNIC-Bsa4 Clean-up**__ Figure 20 The figure above is of the first measured sample nanodrop for pNIC-Bsa4. Figure19 The figure above is of the second measure sample of nanodrop for pNIC-Bsa4 clean-up.

__Analysis:__ The average of the two images is 47.05 ng/uL. A good reading would be around 50 ng/uL. My reading is not as high as I would have hoped or liked it to be, but I will continue with this round of cloning and see how it goes. The next part of lab will be gel checking the sample to ensure that it cut in the right spots. __**PCR Clean-up**__ Figure 18 The figure above is of the first measured sample of nanodrop for PCR clean-up. Figure 17 The figure above is of the second measured sample of nanodrop for PCR clean-up.

__Analysis:__ The average between the two measurements is 64.25 ng/uL. This is a pretty good measurement. The next steps will be to continue with doing pNIC-Bsa4 clean-up.

Very good! -UM

=__Weeks 7 & 8__= __**MidiPrep (2nd trial)**__ Figure 16 The figure above is of the first measured sample of nanodrop (2nd trial).

Figure 15 The figure above is of the second measured sample of nanodrop (2nd trial).

__Analysis:__ This run of midiprep turned out a lot better than the previous one. The average yield here is 58.25ng/uL. This is about the average yield. This time a different pNIC-BSA4 plate was used. The colonies on this plate weren't as close together and they were also a little bigger in size. I believe that the next step would be to cut pNIC-BSA4.

__**MidiPrep (1st trial)**__ Figure 14 The figure above is of the first measured nanodrop image.

Figure 13 The figure above is of the second measured nanodrop image.

__Analysis:__ This run of midiprep did not go the way that it was expected. The average yield of the accepting vectors is 8.25ng/uL. This is not a good yield. There is basically nothing there. One possible reason for this could be that the plate of pNIC-BSA4 that I used had a lot of small colonies. This makes it hard to just get one. I believe that the plate was not a good one because other people who used that plate have had similar results. The next step would be to redo this lab and hopefully get a better yield.

__**PCR^2 (2nd trial)**__ Figure 12 The figure above is of the second trial of PCR^2. This is a 1% agarose gel loaded with 1X TAE buffer. Lane one contains the 100bp ladder. Lane two-five contain samples five-eight respectively. Lane six contains the 1Kb ladder. The 100bp ladder reference is to the left of the gel image.

__Analysis:__ The second trial of PCR^2 went the way that it was expected to go. The samples are all in the right range and they are bright which means that there should be a good yield of the gene. The 1Kb ladder was just run with the samples as a second set of reference. It wasn't needed, I just wanted to make sure that the samples were in the right range. I can now proceed to PCR clean-up.

__**PCR^2 (1st trial)**__ Figure 11 The figure above is of the first trial of PCR^2. This is a 1% agarose gel loaded with 1X TAE buffer. Lane one contains the 100bp ladder. Lane two-five contain samples one-four respectively. The 100bp ladder reference is to the left of the gel image.

__Analysis:__ This trial of PCR^2 did not go as planned. The bands seem to be around the same spot, but they are not bright. Brightness indicates a good yield of the gene. The purpose of doing PCR^2 is to make a lot of the sample, so if the band is not bright, then there is probably not a lot of the sample. This step needs to be redone in order to ensure that there is enough of the sample.

__**Secondary PCR**__

=
Figure 10 The figure above is of the first trial of secondary PCR. This is a 1% agarose gel loaded with 1X TAE buffer. Lane one contains the 100 bp ladder. Lane two contains the primary PCR sample, the working one. Lane three contains the secondary PCR sample. The 100 bp ladder reference is to the left of the gel image.=====

__Analysis:__ It looks like the secondary PCR worked. The primary PCR has the smear which is to be expected. The secondary PCR sample is in the correct range being that the gene has around 700 bp. The sample is also very bright which is good because that means that there is a good amount of the sample. There is a slight band in lane four. Since there is no sample there, there shouldn't be a band. A reasonable explanation for this could be that there was a tear in the gel and some of the secondary PCR sample went into lane four. The gel was also dropped on the floor which is the reason for the rips on the bottom of the gel image. The next step would be to do PCR^2.

__Weeks 5 & 6__
Figure 9 The figure above is of the third trial of Primary PCR. This is a 1% agarose gel loaded with 1XTAE buffer. Lane one contains the 100bp ladder. Lane two contains the Primary PCR sample. The 100bp ladder reference is to the left of the gel image.
 * __Primary PCR (3rd try)__**

__Analysis:__ It looks as thought this trial worked for Primary PCR. This image has a well defined smear. This time I used Brandy's oligo mix because her Primary PCR worked for her the first time. Our results were a little different, but I still believe that mine worked. Now I can move onto Secondary PCR! __**Made LB+KAN agar plates 10/03/13**__ __Analysis:__ When I was using the pipette to dispense the mixture into the petri dishes, a few bubbles made it into the plates. Unfortunately, I underestimated how quickly the mixture would solidify and when I went back to remove the bubbles, they were already solid. __**Primary PCR (2nd try)**__ Figure 8 The figure above is of the 1% agarose gel run with 1XTAE buffer. Lane 1 contains the 100bp ladder and lane 2 contains the primary PCR sample. The 100bl ladder reference is to the left of the gel.

__Analysis__: It looks as though my latest primary PCR run has failed. Im not really sure as to why this is. This run is a lot better than my previous one but the smear needs to be up higher in between the 700 an 800 range. A possible reason for this could be possible contamination. That can cause the smear to be a lot lower. For this run I used Caroline's oligo mix because her primary PCR worked. I thought that if I used hers, I may get the right results. On my next try I will use Brandy's and hopefully it will work. Due to exams over the last couple of weeks I was not able to get as far in lab as I wanted to, but I will be coming in early this week so that I can get on schedule.

*I need week 1 & 2 graded-DSP - Done - Dr. B 092513
=Week 3 & 4= D'Ondria - good work on yoru Week 3 & 4. Move on to retrying Primary PCR and then onto Secondary - Dr. B 092513 __**Primer Tails Came In**__ Figure 7 The figure above is of the forward primers of STP1. They arrived 09/20/2013. Figure 6 The figure above is of the reverse primers for STP1. They came in on 09/20/2013

__Analysis:__ These primers are necessary to do the secondary pcr. Now that these are in, that can be done. __**Primary PCR**__ Figure 6 The figure above is of the 1% agarose gel loaded with 1XTAE buffer. Well one contains the 100bp ladder. Well two contains the primary PCR(1uM oligo mix). The ladder reference it to the left of the gel image.

__Analysis:__ It looks as though this gel run failed. The smear that is supposed to be present in lane two is not present. I can see a small hint of a dark spot but im not sure if that is a hint of am smear or if it just something in the gel. Possible causes for this could be that I didn't run my gel for long enough. An hour is usually a good amount of time but I guess it needed longer. __**Primer Design Tails**__ Upstream: TACTTCCAATCCATG Downstream: TATCCACCTTTACTG Forward primer: TACTTCCAATCC ATGGAAATCTCTCTCCTGACT ___33__ __bp GC Content__ 42.4_% 0 mM Mg2+ Tm ___61.7__ __oC 1.5 mM Mg2+ Tm__ _69 oC 2 mM Mg2+ Tm ___69.6__ __oC__ __4 mM Mg2+ Tm__ _70.6 oC 6 mM Mg2+ Tm ___71__ __oC__ __ Reverse primer: TATAGTTGAATCTGAAGCTGTTTAA CAGTAAAGGTGGATA __ __Reverse complement: TATCCACCTTTACTGTTAAACAGCTTCAGATTCAACTATA __ _40 bp GC Content ___32.5____%__ __0 mM Mg2+ Tm__ _60.3 oC 1.5 mM Mg2+ Tm ___68.5__ __oC 2 mM Mg2+ Tm__ _69 oC 4 mM Mg2+ Tm ___70__ __oC 6 mM Mg2+ Tm__ _70.5 oC

__**Accepting Vector with STP1 insert. 6HIS tag is orange, tail primers are red, start and stop codons are bolded, CDS is underlined.**__ TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATG CACCATCATCATCATCAT TCTTCT GGTGTAGATCTGGGTACCGAGAACCTG TACTTCCAATCC __ **ATG**GAAATCTCTCTCCTG __ __ ACTGACATCGGTCAACGTCGCTCTAATAACCAGGACTTCATCAATCAGTTCGAAAAC __ __ AAGGCCGGTGTTCCGCTCATCATCCTGGCGGACGGCATGGGCGGTCACCGTGCGGG __ __ TAACATTGCGAGCGAAATGACCGTTACCGATCTGGGCTCTGACTGGGCGGAAACCG __ __ ACTTCTCTGAACTGTCTGAAATCCGTGACTGGATGCTCGTTTCTATCGAAACGGAAA __ __ ACCGTAAAATCTACGAACTGGGTCAGTCTGACGACTACAAAGGTATGGGTACCACC __ __ ATCGAAGCGGTTGCGATCGTTGGCGACAACATCATCTTCGCGCACGTTGGTGACTCT __ __ CGTATCGGTATCGTTCGTCAGGGTGAATACCATCTGCTGACTTCCGACCACTCTCTG __ __ GTTAACGAGCTGGTGAAAGCGGGTCAACTGACCGAAGAAGAAGCGGCGTCTCACCC __ __ GCAGAAGAATATCATCACCCAGTCTATTGGCCAGGCGAACCCGGTTGAACCGGACC __ __ TGGGCGTCCACCTGCTGGAAGAAGGTGACTACCTGGTTGTTAACTCTGACGGTCTGA __ __ CCAACATGCTGTCTAACGCGGACATCGCGACCGTTCTGACGCAGGAAAAAACCCTG __ __ GACGACAAAAATCAGGACCTGATCACTCTCGCTAACCATCGTGGTGGTCTGGACAAT __ __ATTACCGTTGCGCTGGTATACGTTGAATCTGAAGCTGTT**T****AA**__ CAGTAAAGGTGGATA CGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCAC CACCACCACTGAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGC TGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAG GGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATTGGCGAATGGGACGCGCCCTGT AGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACT TGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTC GCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGT GCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGG CCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATA GTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGA TTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACA AAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCACTTT TCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATG TATCCGCTCATGAATTAATTCTTAGAAAAACTCATCGAGCATCAAATGAAACTGCAA TTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAA GGAGAAAACTCACCGAGGCAGTTCCATAGGATGGCAAGATCCTGGTATCGGTCTGC GATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAG GTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAA GTTTATGCATTTCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAA ATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAA TACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAATGCAACCGGCGCA GGAACACTGCCAGCGCATCAACAATATTTTCACCTGAATCAGGATATTCTTCTAATA CCTGGAATGCTGTTTTCCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGGAG TACGGATAAAATGCTTGATGGTCGGAAGAGGCATAAATTCCGTCAGCCAGTTTAGTC TGACCATCTCATCTGTAACATCATTGGCAACGCTACCTTTGCCATGTTTCAGAAACAA CTCTGGCGCATCGGGCTTCCCATACAATCGATAGATTGTCGCACCTGATTGCCCGACA TTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCG GCCTAGAGCAAGACGTTTCCCGTTGAATATGGCTCATAACACCCCTTGTATTACTGTT TATGTAAGCAGACAGTTTTATTGTTCATGACCAAAATCCCTTAACGTGAGTTTTCGTT CCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTT TCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTG TTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGC GCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCC AGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAG GCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAAC GACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTC CCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGA GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTT TCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCT ATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTT TGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCC TTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGT GAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCG GTATTTCACACCGCATATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATA GTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGA CACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCT TACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCA TCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGAAGCGA TTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTT AATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCA CTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAAC GAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAA CGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGGACCAGAGAAAAATCAC TCAGGGTCAATGCCAGCGCTTCGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCA GCAGCATCCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGT TTCCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGTCGC AGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGATTCATTCTGC TAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACGACAGGAGCACGATC ATGCGCACCCGTGGGGCCGCCATGCCGGCGATAATGGCCTGCTTCTCGCCGAAACGTT TGGTGGCGGGACCAGTGACGAAGGCTTGAGCGAGGGCGTGCAAGATTCCGAATACCG CAAGCGACAGGCCGATCATCGTCGCGCTCCAGCGAAAGCGGTCCTCGCCGAAAATGA CCCAGAGCGCTGCCGGCACCTGTCCTACGAGTTGCATGATAAAGAAGACAGTCATAA GTGCGGCGACGATAGTCATGCCCCGCGCCCACCGGAAGGAGCTGACTGGGTTGAAGG CTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAAT TGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAA TGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTGGTTTT TCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAG AGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGG TGGTTAACGGCGGGATATAACATGAGCTGTCTTCGGTATCGTCGTATCCCACTACCGA GATATCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGCGC CATCTGATCGTTGGCAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGC ATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCT GAATTTGATTGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGA CAGAACTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGAT GCTCCACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTG TCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACA GCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGC GCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCATC GACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAAT TTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAACGAC TGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATC GCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACG CGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTAC TGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACC GCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCG ACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCGCCGCC GCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGC CTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGCCCG ATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGC CGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGATCCCGCGA AAT Figure 5 The figure above is of the accepting vector being cut by BsaI from the NEBcutter website. The insert for STP1 doesn't get cut by BsaI.
 * The sequence margins are a little off. I couldn't figure out how to copy it correctly from my word document. The order is correct, the margins aren't though. *

__Analysis:__ In order to get melting points that were relatively close base pairs had to be taken out of the forward primer and added to the reverse primer. This was necessary to decrease the chance of messing something up. The GC contents are good because they are below 50%. __**Restriction Enzyme Digest**__ Figure 4 The figure above is of a 1% agarose gel run with 1X TAE buffer. Well one was skipped. In well two there is the 1kb ladder. Lane three contains the uncut plasmid(without blue juice), pGBR22. Lane four contains pGBR22 cut with EcoRI. Lane five contains pGBR22 cut with PvuII. Lane six contains pGBR22 cut with EcoRI+PvuII. Lane seven contains the uncut plasmid (with blue juice), pGBR22. The ladder reference is to the left of the gel picture.

__Analysis:__ I believe that the band didn't appear the way that they should have. There are no bands for sample C, and the other samples seemed to be a little higher than the were expected to be. I had to do another uncut plasmid sample because I forgot to add the blue juice before adding the sample to the well. __**PCR Round One**__ Figure 3 The figure above is of a 1% agarose gel run with 1X TAE buffer. In well one there is 100bp ladder. Lane two contains sample A which is 0.3ng pGBR22. Lane three contains sample B which is 3ng pGBR22. Lane four contains sample C which is 30ng pGBR22. Lane five contains sample D which has no pGBR22. The ladder reference is to the left of the gel picture.

__Analysis:__ I think my first PCR turned out alright. Samples A, B, and C all show up nicely and they are on ladder markers. The only issue is that Sample D shows up. Since there is no plasmid, there should be no line in that lane. Possible reasons for this could be contamination. though I tried to take the proper steps to prevent this, it is possible that contamination still occurred.

=Week 1&2= __**Primer Design**__ __Protein sequence__ 1 MEISLLTDIGQRRSNNQDFINQFENKAGVPLIILADGMGGHRAGNIASEMTVTDLGSDWA 61 ETDFSELSEIRDWMLVSIETENRKIYELGQSDDYKGMGTTIEAVAIVGDNIIFAHVGDSR 121 IGIVRQGEYHLLTSDHSLVNELVKAGQLTEEEAASHPQKNIITQSIGQANPVEPDLGVHL 181 LEEGDYLVVNSDGLTNMLSNADIATVLTQEKTLDDKNQDLITLANHRGGLDNITVALVYV 241 ESEAVX __DNA sequence #1__ 1 ATGGAAATCTCTCTCCTGACGGACATTGGTCAGCGTCGTTCTAACAACCAGGACTTCATC 61 AACCAATTTGAAAACAAAGCGGGCGTGCCGCTCATCATCCTGGCGGACGGCATGGGTGGT 121 CATCGTGCGGGTAATATCGCGTCTGAAATGACTGTAACCGACCTGGGTAGCGACTGGGCG 181 GAAACGGACTTCTCTGAACTGAGCGAAATCCGTGACTGGATGCTGGTCTCTATCGAAACC 241 GAAAACCGTAAAATCTACGAGCTGGGCCAGTCTGACGACTACAAAGGTATGGGTACCACC 301 ATCGAAGCGGTTGCGATCGTTGGTGACAACATCATTTTCGCGCACGTCGGTGACTCTCGT 361 ATCGGCATCGTTCGTCAGGGTGAATACCACCTGCTGACCTCTGACCACTCTCTGGTTAAC 421 GAACTCGTTAAAGCCGGCCAGCTGACCGAAGAGGAGGCTGCAAGCCACCCGCAGAAGAAC 481 ATTATCACCCAATCTATCGGCCAAGCGAACCCGGTTGAACCGGACCTCGGTGTACATCTG 541 CTCGAAGAAGGTGACTACCTCGTTGTAAACTCTGACGGCCTCACGAACATGCTGTCTAAC 601 GCGGACATCGCGACCGTTCTCACGCAGGAAAAAACCCTGGACGACAAAAATCAGGACCTG 661 ATCACCCTCGCGAACCACCGTGGTGGTCTGGACAATATCACCGTTGCGCTGGTTTACGTT 721 GAATCTGAAGCCGTTTAA __18 oligonucleotides to be sequenced__ __**Nanodrop**__ Figure 2 The figure above is of the second measurement of pGBR22. It measures 407.2ng/uL.
 * 1 ATGGAAATCTCTCTCCTGACGGACATTGGTCAGCGTCGTTCTAACAACCAGGACT 55 **
 * 2 TGAGCGGCACGCCCGCTTTGTTTTCAAATTGGTTGATGAAGTCCTGGTTGTTAGAACGAC 60 **
 * 3 GGGCGTGCCGCTCATCATCCTGGCGGACGGCATGGGTGGTCATCGTGCGGGTAATATCGC 60 **
 * 4 CCGCCCAGTCGCTACCCAGGTCGGTTACAGTCATTTCAGACGCGATATTACCCGCACGAT 60 **
 * 5 GGTAGCGACTGGGCGGAAACGGACTTCTCTGAACTGAGCGAAATCCGTGACTGGATGCTG 60 **
 * 6 CCAGCTCGTAGATTTTACGGTTTTCGGTTTCGATAGAGACCAGCATCCAGTCACGGATTT 60 **
 * 7 ACCGTAAAATCTACGAGCTGGGCCAGTCTGACGACTACAAAGGTATGGGTACCACCATCG 60 **
 * 8 GTGCGCGAAAATGATGTTGTCACCAACGATCGCAACCGCTTCGATGGTGGTACCCATACC 60 **
 * 9 ACAACATCATTTTCGCGCACGTCGGTGACTCTCGTATCGGCATCGTTCGTCAGGGTGAAT 60 **
 * 10 AGTTCGTTAACCAGAGAGTGGTCAGAGGTCAGCAGGTGGTATTCACCCTGACGAACGATG 60 **
 * 11 CCACTCTCTGGTTAACGAACTCGTTAAAGCCGGCCAGCTGACCGAAGAGGAGGCTGCAAG 60 **
 * 12 GCTTGGCCGATAGATTGGGTGATAATGTTCTTCTGCGGGTGGCTTGCAGCCTCCTCTTCG 60 **
 * 13 CCCAATCTATCGGCCAAGCGAACCCGGTTGAACCGGACCTCGGTGTACATCTGCTCGAAG 60 **
 * 14 TTCGTGAGGCCGTCAGAGTTTACAACGAGGTAGTCACCTTCTTCGAGCAGATGTACACCG 60 **
 * 15 TCTGACGGCCTCACGAACATGCTGTCTAACGCGGACATCGCGACCGTTCTCACGCAGGAA 60 **
 * 16 CGCGAGGGTGATCAGGTCCTGATTTTTGTCGTCCAGGGTTTTTTCCTGCGTGAGAACGGT 60 **
 * 17 ACCTGATCACCCTCGCGAACCACCGTGGTGGTCTGGACAATATCACCGTTGCGCTGGTTT 60 **
 * 18 TTAAACGGCTTCAGATTCAACGTAAACCAGCGCAACGGTG 40 **

Figure 1 The figure above is of the first measurement of pGBR22. It measures 410.6ng/uL

__Analysis:__ The purpose of this lab was to determine the concentration of the plasmid using the nanodrop software and the spectrophotometer. The only possible error is that the 260/230 values aren’t that close to 2.1. They should be near that but for some reason they aren’t. The next steps after this lab would be to possibly do a third trial to see if the 260/230 values could be closer to 2.1 which is the desired value.