Team:Brown/Notebook Meetings/7-13-09

'''iGEM Meeting Minutes '''

'''July 13, 2009 '''

9:00 AM 

'''SFH 218 '''


 * 9:07AM: Team 1 update
 * Sent out EV131 for sampling last week, the gene is in there
 * EcoR1 and BamH1 digest is not working
 * There is a mistake in the primers… They added an inadvertent ATG in the prefix. They designed new ones and will run them by Gary.
 * Now its just getting the appropriate primers with appropriate restriction sites, they will order T7 and T3 primers
 * Gary suggests placing the enzymes in Styrofoam to keep it stable
 * After digest, ligate into pGEM, then into pNoTat


 * 9:22: Team 2 update
 * Last week they sent their project proposal to Looger
 * Received secretion tag from Geneart, made a nice lawn, miniprepped
 * Digest is hard because it is difficult to resolve small DNA
 * Run in a higher percentage gel (1.5% - 2%)
 * Received PCR primers for Agr operon, however PCR will not work
 * What do you do now? It would be a good control to use the previous S. epi genomic DNA as a PCR reaction that works.
 * When doing digest on PCR product, incubate overnight.


 * 9:40: Team 3 update
 * Current goal is to get all genes functional in RU1012 strain.
 * Last week worked on OmpC over TetA, did 2 digests, 1st one showed a good digest but wrong enzymes. 2nd digest with correct enzymes did not have any cuts.
 * We will be doing the digest again.
 * Eli suggests using another gene besides TetA that will allow weakly expressing cells to survive
 * Mutagenic primers designed
 * Small molecule library will be used to screen


 * 10:00: Gary will be out Thursday until all next week.