1191 pCDNA3.1-Dest2

5/22/2012

digest pCDNA3.1-Dest 1190 with Not1 + BamH1 (Lane #3).
cut out vector (not the small fragment), purify with column, elute with 20 ul water.
Blunt end:
Add 2.5 ul buffer 4, 1 ul of 10 mM dNTPs.
Add 1 unit (0.3 ul of 3 u/ul) T4 DNA Polymerase
incubate 15 minutes at 12°C.
Purify:
Add gel extraction buffer and purify with column.
ligate and transform ccdb survival.

5/24/2012

Cloning of 1191 pCDNA3.1-Dest2

Purpose: Cloning of1191 pCDNA3.1-Dest2

Date: 2012-6-28---2012-7-5

Notebook:0

Page:

Clone ID:1191

Experimenters: yingying

Results

cloning

ligate to annealed oligos
GGCCGA ACGCCGATATTGAAGGTATT c
ct TGCGGCTATAACTTCCATAA gCTAG

PfuMBP-seq-F
GGCCGA ACGCCGATATTGAAGGTATT c
GW-fix-B
GATCgAATACCTTCAATATCGGCGTtc

		band 4
		1238pCDNA3.1-Dest 1238
enzymes	Not1 + BamH1        .

Colony Screening

Date: 2012-7-5

Make miniprep and digest with Kpn1+Xho1

Result intepretation/Summary

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