2350 pFastbac-GP64
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Cloning of 2350 pFastbac-GP64
[edit]Purpose: Cloning of2350 pFastbac-GP64==
Date: 2014-9----
Notebook:0
Page:
Clone ID:2350
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from 2351 pEC2-GP64
Fragment 1
2350-GP64-Xho1-F tacgac ctcgag ATGGTAAGCGCTATTGTTTTATATG
2350-GP64-R1 (gaaaccgtaatagacaatattaa gtac) gtacttaatattgtctattacggtttc
Fragment 2
2350-GP64-Xho1-F tacgac ctcgag ATGGTAAGCGCTATTGTTTTATATG
2350-GP64-R2 (gaaaccgtaatagacaatattaa) ttaatattgtctattacggtttc
purify two fragments in one tube, heat to 95 oC, cool slowly to RT then digest with Xho1 (without Kpn1)
ligate to Xho1 + Kpn1 digested vector
Colony Screening
[edit]Date: 2014_8
Make miniprep, use Xho1 + Kpn1 to find positive clone
