IGEM:MIT/2007/Notebook/2007-7-18

Agenda

1. Transform tk's competent cells with yesterday's 3A assembly results (F2620+B0034 and CPX insert into vector pSB1AC3)
2. Make LB+Cm and LB+Cm+Kan plates
3. Liquid cultures of eight tubes made and put in warm room -- for making glycerol stocks

• • 2 tubes of DB3.1 (Kan+)
  • 2 tubes of transformed E1010 from resistance test plate (Kan+)
  • 2 tubes of transformed F2620 from resistance test plate (Amp+)
  • 2 tubes of transformed B0034 from resistance test plate (Amp+)
  • All should be taken out tomorrow morning and used to make glycerol stocks

1. Meeting with TK and Drew

Transformed tk's competent cells

• Aliquoted one of the original vials into eight eppendorfs, 100µL each.
• Did 3 sets of transformations:
  • 2 of yesterday's 3A assembly product (plasmid contains: F2620+B0034+CPX in pSB1AC3 backbone)
  • 1 negative control (pSB1AC3 digested with EcoRI and PstI)
• Followed our Transformation protocol (tk's modified protocol)

• Transformations were put into 37C room at 11:30 AM and incubated for 2 hrs

Poured Plates, again

• Used stringent working concentrations:
  • 10µg of Kanamycin per mL of LB
  • 25µg of Chloramphenicol per mL of LB
• Poured 5 LB+Cm+Kan plates
• Poured 29 LB+Cm plates
• Cm plates have green labels