1300 pET-6H-KLPVM-NLS
Cloning of 1300 pET-6H-KLPVM-NLS
[edit]Purpose: Cloning of 1300 pET-6H-KLPVM-NLS
Date: August 13, 2012
Notebook:
Page:
Clone ID: 1300
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: August 13, 2012
Anneal two oligos
His-KLPVM-NLS-T
cATGGCACATCATCATCACCATCATAAACTGCCGGTTATGCCGAAAAAAAAACGCAAAGTTca
His-KLPVM-NLS-B
tatgAACTTTGCGTTTTTTTTTCGGCATAACCGGCAGTTTATGATGGTGATGATGATGTGcGc
Ligate and transform to Nco1 +Nde1 digested 1128 pDB.His.PfuMBP-(tev), see lane 2
Date: August 15, 2012
Anneal two oligos
His-KLPVM-NLS-T
cATGGCACATCATCATCACCATCATAAACTGCCGGTTATGCCGAAAAAAAAACGCAAAGTTca
His-KLPVM-NLS-B
tatgAACTTTGCGTTTTTTTTTCGGCATAACCGGCAGTTTATGATGGTGATGATGATGTGcGc
Ligate and transform to Nco1 +Nde1 digested pDB.His.MBP, V14, see lane 2
Transform
Colony Screening
[edit]Date: August 17, 2012
Digested MiniPrep with RE Nde1 and Nco1
Very faint insert band at expected annealed origo size ~180 bp
Result intepretation/Summary
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Conclusion
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