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

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(Agenda)
(Agenda)
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#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
#Liquid cultures of eight tubes made and put in warm room -- for making glycerol stocks
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**2 tubes of DB3.1 (Kan+)
+
##2 tubes of DB3.1 (Kan+)
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**2 tubes of transformed E1010 from resistance test plate (Kan+)
+
##2 tubes of transformed E1010 from resistance test plate (Kan+)
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**2 tubes of transformed F2620 from resistance test plate (Amp+)
+
##2 tubes of transformed F2620 from resistance test plate (Amp+)
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**2 tubes of transformed B0034 from resistance test plate (Amp+)
+
##2 tubes of transformed B0034 from resistance test plate (Amp+)
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**All should be taken out tomorrow morning and used to make glycerol stocks
+
##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 12:48, 18 July 2007

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
    1. 2 tubes of DB3.1 (Kan+)
    2. 2 tubes of transformed E1010 from resistance test plate (Kan+)
    3. 2 tubes of transformed F2620 from resistance test plate (Amp+)
    4. 2 tubes of transformed B0034 from resistance test plate (Amp+)
    5. All should be taken out tomorrow morning and used to make glycerol stocks
  4. 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
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