Transformed colony culture and VR agarose gel extraction #

Culture preparation #

Retrieved the plates from the transformation I did yesterday from the 37C room around 11am. All transformations were successful except for VR-7 and VR-13. Additionally the negative control showed no growth while there were several colonies present on the positive control plate.

I transferred several colonies from each plate into 8ml of LB + amp and placed tubes onto the shaker in the 37C room at 200 rpm to grow overnight.

Replated VR-7 and 13 #

Since I saved the transformed bacteria from yesterday I figured I would just replated the failed transformation just to be sure it was in fact the transformation that failed and not some other weird plating fluke. Placed those plates in the 37C room at around 1pm to grow overnight.

VR-29, 30 and 31 agarose gel extractions #

Used freeze and squeeze method to extract small bands (VR fragments) from VR-29, 30, and 31 PCR / BglII digestion products I set up yesterday. Nanodrop results from the extractions are below. VR-29 and 30 were ran in two lanes while VR-31 was ran in three due to the larger volume of the sample after BglII digestion. Approximate sample volume was 50ul.

CSV version of the table is also available here.After nanodroping I combined samples (1 sample per insert in the end) and placed into the VR-inserts 2 box in the kitchen freezer.