2273 pHCMC04-Cre
Cloning of 2273 pHCMC04-Cre
[edit]Purpose: Cloning of2273 pHCMC04-Cre==
Date: 2014-7----
Notebook:0
Page:
Clone ID:2273
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from 298
6His-BamH1-F B4 tactaggatccatgggccatcaccatcaccatcac
Cre-Xma1-R B5 tgatacccgggCTCGAGCTAATCGCCATCTTCCAGCAG
digest with BamH1+ Xma1, clone to 2271 pHCMC04
Colony Screening
[edit]Date: 2014_7
Make miniprep, use XhoI to find positive clone
use BamH1+ Xma1 to verify
sequenced, not right , there is a BamHI site in cre sequence
Cloning of 2273 pHCMC04-Cre
[edit]Purpose: Cloning of2273new pHCMC04-Cre==
Date: 2014-7----
Notebook:0
Page:
Clone ID:2273
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from 298
2 fragments
1st fragment( the lower band)
6His-BamH1-F tactaggatccatgggccatcaccatcaccatcac
Cre-dBamH1-R CTTTTCGGATaCCGCCGCATAAC
2nd fragment
Cre-dBamH1-F GTTATGCGGCGGtATCCGAAAAG
Cre-Xma1-R tgatacccgggCTCGAGCTAATCGCCATCTTCCAGCAG
join 2 fragment
6His-BamH1-F tactaggatccatgggccatcaccatcaccatcac
Cre-Xma1-R tgatacccgggCTCGAGCTAATCGCCATCTTCCAGCAG
digest with BamH1+ Xma1, clone to 2271 pHCMC04
Colony Screening
[edit]Date: 2014_7
Make miniprep, use XhoI to find positive clone
use BamH1+ Xma1 to verify
Cloning of 2273 pHCMC04-Cre
[edit]Purpose: Cloning of2273 pHCMC04-Cre==
Date: 2014-8----
Notebook:0
Page:
Clone ID:2273
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from 298
2 fragments
1st fragment
6His-BamH1-F tactaggatccatgggccatcaccatcaccatcac
2273-Cre-dBamH1-R2 CTTTTCGGATaCGCCGCATAAC
2nd fragment
2273-Cre-dBamH1-F2 GTTATGCGGCGtATCCGAAAAG
Cre-Xma1-R tgatacccgggCTCGAGCTAATCGCCATCTTCCAGCAG
join 2 fragment
6His-BamH1-F tactaggatccatgggccatcaccatcaccatcac
Cre-Xma1-R tgatacccgggCTCGAGCTAATCGCCATCTTCCAGCAG
digest with BamH1+ Xma1, clone to 2271 pHCMC04
Colony Screening
[edit]Date: 2014_8
Make miniprep, use XhoI to find positive clone
use BamH1+ Xma1 to verify
