Berk2006-LocksAndKeysTeam

KAD_06/19/06

• Ran analytical gel on PCR products (A (the 994 key3 library) and B (J01093)) made 06/17/06. Visible bands present for each sample confirmed success of PCR.

EIPCR of 994 key3 library

• Digested and recircularized 994 key3 library created by PCR.
• Transformed 994 key3 library into GeneHogs and TGI with pSB3C6-J01122 by electroboration. Plated products.
• Cotransformed J01122 and J01129 by heat shock. Plated product.

KAD_06/22/06
1. We grew cultures of pSB3C6-J01022, pSB3C6-J01122-A, pSB3C6-J01093, and to saturdation.
2. Put samples in tecan
3. Measured the fluorescence:

• pSB3C6-J01122; at saturation: 20, at midlog: 31
• pSB3C6-J01022; at saturation: 2002, at midlog: 267
• pSB3C6-J01093: at saturation: 24, at midlog: 19
• pSB3C6-J01122/pSBIA2-J01129; at saturation: 2484, at midlog: 311

We did this experiment to test our locks and our keys. In order to do this, we had to compare results with this test of cells with RFP only (pSB3C6-J01122), the lock only (pSB3C6-J01122), both the lock and the key (pSB3C6-J01122/pSBIA2-J01129), and cells containing nothing (pSB3C6-J01093).