2061 pEC-ExcellScript-II
Cloning of 2061 pEC-ExcellScript-II
[edit]Purpose: Cloning of2061 pEC-ExcellScript-II==
Date: 2014-2----
Notebook:0
Page:
Clone ID:2061
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR
PCR two fragments:
Fragment 1
ssIII-Nde1-F1
ctagcCATATGACCCTAAATATAGAAGATGAGCATCGGCTAC
ssIII-M289L-R4
(GAGGCCAGAAAA GAGACTGTG cTG GGG CAGCCTACTCCGAAG ACCCCTCGACAACTAAG)
CTTAGTTGTCGAGGGGTCTTCGGAGTAGGCTGCCCCAgCACAGTCTCTTTTCTGGCCTC
Fragment 2
ssIII-V223H-F4
CAGACTTCCGGATCCAGCACCCAGACTTGATCCTGCTACAGTACcatGATGACTTACTGCTG
ssIII-Xho1-R8
(TCACAGAGAATCCAGACACCTCTACCCTCCTCATAGAAAATTCATCACCGAATTCCCGCTTAATTAATCTCGAGctag)
ctagCTCGAGATTAATTAAGCGGGAATTCGGTGATGAATTTTCTATGAGGAGGGTAGAGGTGTCTGGATTCTCTGTGA
2. Join two fragments using
ssIII-Nde1-F1 + ssIII-Xho1-R8
3. Digest with Nde1 + Xho1, clone into pEC
Colony Screening
[edit]Date: 2013-10-23
Make miniprep, digest with Nde1 + Xho1+BamHi
