1504 pT7A3-T4DNA-Pol-Exo-
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Cloning of 1504 pT7A3-T4DNA-Pol-Exo-
[edit]Purpose: Cloning of1504 pT7A3-T4DNA-Pol-Exo-==
Date: 2014-9----
Notebook:0
Page:
Clone ID:1504
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR using 1765 pET28b-T4DNAPol as template
Fragment 1
T4DNAPol-1-1 CGACTAGCTAGCAAAGAATTTTATATCTCTATTGAAACAGTCGG
T4-p43D219A-R catgatatacggaacgGcaaacccctcaatattc
Fragment 2
T4-p43D219A-F gaatattgaggggttt gCc gttccgtatatcatg
T4DNAPol-3-24 CGATCGGCGGCCGCTCAGCCAAACAGGAAGTCTAACGAAGCTTTTTCTTCATA
join two fragments by PCR
digest with NheI+NotI
Colony Screening
[edit]Date: 2014_8
Make miniprep, use HindIII to find positive clone
