IGEM:IMPERIAL/2006/LabCalendar/2006-8-8

Electroporation of Ligations from yesterday

 * Electroporated 12D->2H (3rd attempt...should be successful??? thrid time's a charm?) and 12D->2H B (the big part from the gel)
 * Plates are in the incubator, needs to be minipreped tomorrow to check...and probably PCR to correlate.

Maxiprep of Ligations from yesterday

 * Results of maxiprep of ligations A, M and Z may be dubious. Should repeat maxiprep for assurance.
 * Maxiprep of ligations L, R and V should be fine.

To ensure we can re-maxiprep tomorrow we need to culture up more bacteria by the end of today.

Deepti and Jo h nny cultured up A with the last of the LB so it should be ready to maxiprep for tomorrow. The other ligations have a good chance of having been maxipreped successfuly so we can decide what to do tomorrow (LBs arriving tomorrow anyway). Below is a picture of the gels showing results of both, minipreps and ligations yesterday and today. Still have to put up the riboswitch gels. From top left, clockwise: Gel 1: Miniprep samples A-I (left to right).Only A considered to be successful and maxi-preped. Odd results assumed to be due to contamination at the gel purification stage. Gel 2: Miniprep samples R-AA (right to left). All assumed to be successful hence samples chosen at random to be maxi-preped. R, V and Z. Gel 3: PCR samples R-AA (left to right). Samples R, S, T and U appear to be successful. Samples for maxi-preps chosen this morning before results of PCR. Sample R appears to be successful inboth PCR and mini-prep analysis. Gel 4: PCR samples A-I (left to right). Sample A appears to be successful in accordance with mini-prep results. Large number of false positives attributed to contamination in earlier step.

Cre part

 * didn't work
 * re-running PCR overnight