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Team:Brown/Notebook Meetings/7-13-09
From 2009.igem.org
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.
