Team:Imperial College London/Wetlab/Calendar/22-09

=22/09= Dave – PCR the SLIC primers for the RCSB parts (ultimately to SLIC construct 13): PcstA + RBS by PCR out from construct 1

 PCR of RcsB and the CRP-RBS (C6) BioBricks. Program 42 used on PCR machine. C6: 50, 52, 54 degrees RcsB: 51, 53, 55 Run on a gel to view. (M, C6 50, 53, 54, RcsB 51, 53, 55) Next step is to ligate to GFP BioBrick in vector Restriction Digest Assay PCR Products from the reaction kept in iGEM DNA box as no space in PCR products box. in jeremy freezer Overnight digest done with 1ul DNA to 1ul enzyme. Needs running on a gel, am tomorrow. Depending on results, can recalculate the ratios for the next digest. Used Dam+ and Dam- template from fridge. contents of RD: 6ul Distilled Water 1ul Genome prep DNA undiluted 1ul BSA (x10) 1ul B2 (x10) 1ul Enzymes Note, did not use the buffers enclosed with the enzymes, not sure if this matters.



Dineka – Extracting DNA from the registry for all controls (5 experiments) + Minis (CRP+RBS+AK3) and their respective restriction digests.

Kun + Charles – Plate OD calibration by today + HVD-GFP (preliminary set of results but without proper controls) + Autoinduction media testing