2254 pT7A3-wtSM2
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Cloning of 2254 pT7A3-wtSM2
[edit]Purpose: Cloning of2254 pT7A3-wtSM2==
Date: 2014-7--2014/8/1--
Notebook:0
Page:
Clone ID:2254
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR using new oligos, 1680 pCold-GST-wtSM2 as template
1 ul of 10 uM SM2-signal-Nde1-F1 agctacatATGCGCTTTAACAACAAGAATGTTGGCCTTGGCGCCCTGCTGTTCGCCGCGCAG
1 ul of 1 uM SM2-signal-F2 GCCCTGCTGTTCGCCGCGCAGGCGTCGGCCgacacgctcgaatccatcgac
1 ul of 10 uM WtSM2-Xho1-R (gagttgatgggctgcaaaaac taa ctcgag ctagca) tgctagctcgagttagtttttgcagcccatcaactc
digest with Nde1 + Xho1, clone to 1727 pT7A30
Colony Screening
[edit]Date: 2014_8_1
Make miniprep, use Nde1 + XhoI to find positive clone
digest with NdeI+EcoRV, can get a 500bp band
