September/4 September 2009

1.Colony check sample No. (no. of colonies) No.5              0 No.9             100~ No.18             3 No.20             0 No.21             1 No.22             0 No.23             0 No.25             0 No.26             0 No.9 ,18, 26 colony were streaked on plate for quick ligation check.

2.ligation No.9-I,V, 18-I,V , 24-I,V , 27-I,V gel cut ↓ purification ↓ ligation 2hr

3.transformation Transform No.20, 22, 23, 25, 9, 18, 24, 27 and incubate at 37℃ overnight.

4.mini prep Mini prep and nano drop check of above DNA.(1-14L 1-15L 2-14L 1-16I 1-1K 1-15J 1-14N No.21)

sample No. conc. 1-14L      14.3 ng/uL 1-15L       7.1 ng/ul 2-14L       9.2 ng/uL 1-16I      11.5 ng/uL 1-1K       11.1 ng/uL 1-15J       7.6 ng/uL 1-14N      15.4 ng/uL No.21      19.9 ng/uL

5.digestion and ligation digestion with restriction enzyme

K204005

↓ 37°C, 2hr ↓ gel electrophoresis No.5-I,V gel cut purification ligation O/N

6.Make LB with agar and Amp plates.

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