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.