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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 Johnny 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