Newcastle University Wetlab/4 August 2008

Monday 4th August

 * All 5 of us (Megan, Mark, Nina, Ria and Jess) went into the lab today to decide on a plan of action for the weeks to come.


 * We used Agarose Gel Electrophoresis to check for the presence of plasmids (pGFPrrnB - our plasmid with the gfp gene, and pJWV021 - with the mCherry gene). Our gel was inconclusive, so we decided to re-run it the following day to confirm our results.


 * In the afternoon we attempted to transform TOP10 competent E. coli cells with DNA from the registry (from source plate 1010 well 4A and 1018 7A). See Transforming into DH5α or TOP10 E. coli


 * We followed the extraction method outlined in the green folder and for each DNA spot we plated two agar dishes: one with a larger volume of DNA (4μl) and one with a smaller volume (2μl) to make colony counting easier. (See Making Agar Plates.) We plated the following and incubated at 37˚C overnight:

Plate 1: +ve control (isolated plasmid plus TOP10 cells)

Plate 2: -ve control (TOP10 cells only, no plasmid)

Plate 3: Large 1010 4A

Plate 4: Small 1010 4A

Plate 5: Large 1018 7A

Plate 6: Small 1018 7A