IGEM:MIT/2007/Notebook/2007-7-18: Difference between revisions
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#Transform tk's competent cells with yesterday's 3A assembly results (F2620+B0034 and CPX insert into vector pSB1AC3) | #Transform tk's competent cells with yesterday's 3A assembly results (F2620+B0034 and CPX insert into vector pSB1AC3) | ||
#Make LB+Cm and LB+Cm+Kan plates | #Make LB+Cm and LB+Cm+Kan plates | ||
#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 | |||
#Meeting with TK and Drew | #Meeting with TK and Drew | ||
Revision as of 09:47, 18 July 2007
Agenda
- Transform tk's competent cells with yesterday's 3A assembly results (F2620+B0034 and CPX insert into vector pSB1AC3)
- Make LB+Cm and LB+Cm+Kan plates
- 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
- 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