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)
1 200 uL 0.2 59 42 101
2 100 uL 0.1 17 16 33
3 50 uL 0.05 3 3 6(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)
A1 Transformed LB-amp + X-gal 101 TNTC
A2 Transformed LB-amp + X-gal 102 TNTC
A3 Transformed LB-amp + X-gal 103 233
B1 Transformed LB only 104 TNTC
B2 Transformed LB only 105 196
B3 Transformed LB only 106 52
C1 Untransformed LB-amp + X-gal 101 0
D1 Untransformed LB only 104 TNTC
D2 Untransformed LB only 105 278
D3 Untransformed LB only 106 77
E1 Original (Incubated) LB only 107 9 (ESPC)
E2 Original (Incubated) LB only 108 0
F1 Original (Pre- Incubation) LB only 106 TNTC
F2 Original (Pre- Incubation) LB only 107 44
OC1 Original (Pre-incubation) LB 106 41
OC2 Original (Pre-incubation) LB 107 4
A1 Transformed LB-amp + Xgal 10 TNTC
A2 Transformed LB-amp + Xgal 102 132
A3 Transformed LB-amp + Xgal 103 10
B1 Transformed LB 105 TNTC
B2 Transformed LB 106 134
B3 Transformed LB 107 22
C Untransformed LB-amp + Xgal 10 0
D1 Untransformed LB 105 TNTC
D2 Untransformed LB 106 38
E1 Original (incubation) LB 106 113
E2 Original (incubation) LB 107 9


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.