2300 pEC-wtSM2-b
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Cloning of 2300 pEC-wtSM2-b
[edit]Purpose: Cloning of2300 pEC-wtSM2-b==
Date: 2014-8----
Notebook:0
Page:
Clone ID:2300
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR using new oligos, 1680 pCold-GST-wtSM2 as template
1 ul of 10 uM SM2-signal-Xba1-F1
tagcTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATAATGCGCTTTAACAACAAGAATGTTGGCCTTG
1 ul of 1 uM SM2-signal-Nde1-F2
ACAAGAATGTTGGCCTTGGCGCCCTGCTGTTCGCCGCGCAGGCGTCGGCCgaccatatgacgctcgaatccatcgac
old oligo
1 ul of 10 uM WtSM2-Xho1-R
(gagttgatgggctgcaaaaac taa ctcgag ctagca)
tgctagctcgagttagtttttgcagcccatcaactc
digest with Xba1 + Xho1, clone to2015
Colony Screening
[edit]Date: 2014_8_
Make miniprep, useXba1 + Xho1 to find positive clone