2409 pT7A3-Tat-dCre-(Tev)-Tat-NLS-iFLP-6H
Cloning of2409 pT7A3-Tat-dCre-(Tev)-Tat-NLS-iFLP-6H
[edit]Purpose: Cloning of2409 pT7A3-Tat-dCre-(Tev)-Tat-NLS-iFLP-6H==
Date: 2015_1
Notebook:0
Page:
Clone ID:2409
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from 298 pTriEx-HTNC
Fragment 1 2409-Cre-Nco1-F tataa ccatgggccatcaccatcaccatcacggcatg
2409-E129R-Q133H-R gaacgctagagcAtgttttgcacgGCGaccggcatcaac
Fragment 2
2409-E129R-Q133H-F gttgatgccggt CGC cgtgcaaaa caT gctctagcgttc
2378-Cre-TEV-Nde1-R (gcctgctggaagatggcgatGGTACCggggatgacgatgacaaggaaaacctttacttccagggcCAtatgaacac) gtgttcataTGgccctggaagtaaaggttttccttgtcatcgtcatccccGGTACCatcgccatcttccagcaggc
Join
2409-Cre-Nco1-F tataa ccatgggccatcaccatcaccatcacggcatg
2378-Cre-TEV-Nde1-R (gcctgctggaagatggcgatGGTACCggggatgacgatgacaaggaaaacctttacttccagggcCAtatgaacac) gtgttcataTGgccctggaagtaaaggttttccttgtcatcgtcatccccGGTACCatcgccatcttccagcaggc
digest with Nco1 + Nde1
Colony Screening
[edit]Date: 2015_1
Make miniprep, use HindIII + Nde1 to find positive clone