User:Mark X. Ling/Notebook/Micb 323/2009/01/27

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Lab 3: cell transformation

Part 1: Heat-shock ligation from previous ligation product

  • time of mix(on ice):1:13 - 1:45
  • Time put in shaking water bath:1:55 - 2:55
Tube Volume plated Final Plated Dilution Blue colonies (cfu) White colonies (cfu) Total colonies (cfu)
1200 uL0.25942101
2100 uL0.1171633
350 uL0.05336(ESPC)

Part 2: comparison of Heat-shock ligation and electroporation

Cacl2/Heat shock ligation (student A)

  • By Crystal Lee

Electroporation (Student B)

  • Time on incubation:2:30 - 3:30
  • original on incubation: 2:40 - 3:48
  • Notes:
    • the Electroporated samples were not shaking for unknown amount of time.
    • Original were incubated for 1 hr and 8 minutes.

  • plate 0.1mL therefore undiluted = 10^-1
Tube Cell plated Plate content Final Plated Dilution Factor Colony Count (cfu)
A1TransformedLB-amp + X-gal101TNTC
A2TransformedLB-amp + X-gal102TNTC
A3TransformedLB-amp + X-gal103233
B1TransformedLB only104TNTC
B2TransformedLB only105196
B3TransformedLB only10652
C1UntransformedLB-amp + X-gal1010
D1UntransformedLB only104TNTC
D2UntransformedLB only105278
D3UntransformedLB only10677
E1Original (Incubated)LB only1079 (ESPC)
E2Original (Incubated)LB only1080
F1Original (Pre- Incubation) LB only106TNTC
F2Original (Pre- Incubation)LB only10744
OC1Original (Pre-incubation)LB10641
OC2Original (Pre-incubation)LB1074
A1TransformedLB-amp + Xgal10TNTC
A2TransformedLB-amp + Xgal102132
A3TransformedLB-amp + Xgal10310
CUntransformedLB-amp + Xgal100
E1Original (incubation)LB106113
E2Original (incubation)LB1079

Post Lab Comments

  • transformation by electroporation is better: was faster to use
    • after seeing the colonies, it seems to be better method also.
  • Plating went smoothly
    • there were inconsistency in # of colonies.
    • may not have let the plate dry for long enough: there were lines of colonies in numerous plate.

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