2315 pEC2-TEV
Cloning of 2315 pEC2-TEV
[edit]Purpose: Cloning of2315 pEC2-TEV==
Date: 2014-9----
Notebook:0
Page:
Clone ID:2315
Experimenters: shanyi
Results
[edit]CLONING
[edit]use the following oligos and 432 pMBP-7xHis-TEV (pMHTDelta238) as template
LysN-Tev-2
Sequence ATCGCCTCTAGATCCCTATCAGTGATAGAGATTG
LysN-Tev-3
Sequence TGTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGATCT
(AGATCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACA)
LysN-Tev-4
Sequence CAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACCGAATTGATTTAAGGAG
LysN-Tev-5
Sequence CTTCCAGCCCGGCACCCATGGATCTTACCTCCTTAAATCAATTCGGTCAGT
(ACTGACCGAATTGATTTAAGGAGGTAAGATCCATGGGTGCCGGGCTGGAAG)
1 ul of 10 uM of
PlateName 2012_3_27
Pos D08
SequenceName LysN-Tev-seq-F
Sequence GTCGATGGATCCCTATTCATTCATGAGTTGAGTC
Year 2012
Date 3/30/2012
gtcgatggatccCTATTCATTCATGAGTTGAGTC
(GACTCAACTCATGAATGAATAGggatcc atcgac)
LysN-Tev-2-->> TCTAGATCCCTATCAGTGATAGAGATTG CAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACCGAATTGATTTAAGGAG LysN-Tev-4-->>
promoter TCTAGATCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACCGAATTGATTTAAGGAGGTAAgATCC TEV--> ATGGGTGCCGGGCTGGAAG
(AGATCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACA) <<--LysN-Tev-3
ACTGACCGAATTGATTTAAGGAGGTAAGATCC ATGGGTGCCGGGCTGGAAG <<--LysN-Tev-5
BamH1 + Xba1
Colony Screening
[edit]Date: 2014_9
Make miniprep, use BamH1 + Xba1 to find positive clone
digest with NdeI+Hindiii to verify
