2255 pT7A3-wtSM2-b
Cloning of 2255 pT7A3-wtSM2-b
[edit]Purpose: Cloning of2255 pT7A3-wtSM2-b==
Date: 2014-7--2014/8/1--
Notebook:0
Page:
Clone ID:2255
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
1 ul of 10 uM WtSM2-Xho1-R (gagttgatgggctgcaaaaac taa ctcgag ctagca) tgctagctcgagttagtttttgcagcccatcaactc
digest with Xba1 + Xho1, clone to 1727 pT7A30
Colony Screening
[edit]Date: 2014_8_1
Make miniprep, use Xba1 + Xho1 to find positive clone
