Checking Addgene plasmids
- We received pMevT and pMBI (for iGEM/Matt C's stuff) earlier this week
- I will test that they grow (today) and produce the expected plasmid map (Mon or Tues)
- I'll streak for isolation, move the plates to the fridge tomorrow, and start O/Ns either Sunday or Monday.
- 37°C on agar plates over the weekend w/ appropriate antibiotic
- Tet stocks were bad, see below
Plasmid maps (for Monday/Tuesday):
- pMBI: Given a rough plasmid map (removed ispA from pMBIS), PstI should cut 2x, yield 5.8, 3.9kb bands.
- pMevT: EcoRV should yield 2 bands, 7.1 & 1.5kb.
Making tetracycline stocks
- Old stocks were insoluble, made a couple years ago
- It's a powder in the back of the bottom of the -20°C
- Dissolved in 190 proof EtOH to 5 mg/mL 1 mL stocks and put in the -20°C
- Tet selective concentration is 10 μg/mL, so the stocks are 500x (vs normal 1000x)