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.