Difference between revisions of "Team:Stony Brook/Notebook/Week8"

Week 8

7/17:

    • Troubleshooting of gel extraction. No high yields up to this point which required repeating a lot of miniprepping and restriction enzyme digestion

    • Learned about gel electroelution as a potential solution instead of gel extraction.

    • Started the gel electroelution process

    • Digested the rest of miniprepped samples from (7/11) and ran on a gel, which is necessary for both gel electroelution and gel extraction

7/18:

    • Finished gel electroelution and nanodropped samples

    • We tried to nanodrop all samples again, but got values that were the same as the first attempt.

    • The results from the nanodrop did not make sense, as the concentration of DNA for each sample exceeded the amount that we digested. We decided to try the regular gel extraction protocol again.

    • Ligation of LnqZA U into MBP, GST, and SUMO vectors

7/19:

    • Received a donation from Truetox Laboratories today!

    • Finish compiling S. Aureus protocols in order to receive BSL2 clearance for our laboratory.

    • Gel extracted the rest of the samples, since gel electroelution failed

7/20:

    • Ligations of MBP vector and LnqZA T, LnqZA U, LnqZE T, LnqZQ, Aureocin A53, and Epidermicin NI01

    • Digestions:

    • Ran on 1.8% agarose gel

    • Gel extracted and nanodropped Gel Extracts. Much better yields, 260/280 values more mixed

7/21:

    • Digestions:

    • Ran on 1.8% agarose gel

    • Nanodrop of Gel Extractions

    • Make liquid cultures of GST, MBP, and SUMO vectors and LnqZA T, LnqZA U, and LnqZE T inserts.

    • Transformation of individual bacteriocins into BL21 E. coli

    • Transformation of LnqZE U into DH5α E. coli</p>