2351 pEC2-GP64
Cloning of 2351 pEC2-GP64
[edit]Purpose: Cloning of2351 pEC2-GP64==
Date: 2014-9----
Notebook:0
Page:
Clone ID:2351
Experimenters: shanyi
Results
[edit]CLONING
[edit]gene synthesis the first fragment
the following fragments should be amplified by PCR from DH10Bac Bacmid
Fragment 2
2351-GP64-F gcggagcactgcaacgcgcaaatg
2351-GP64_dBamH1_1-R gtgtgttgggatcGAGtgagccgccgttg
Fragment 3
2351-GP64_dBamH1_1-F caacggcggctcaCTCgatcccaacacac
2351-GP64_dXho1-R cttggcactcCagcctgctgtac
Fragment 4
2351-GP64_dXho1-F gtacagcaggctGgagtgccaag
2351-GP64_dNot1-R caagtgggTggccgcttcttg
Fragment 5
2351-GP64_dNot1-F caagaagcggccAcccacttg
2351-GP64_dBamH1_2-R gttgccgatggtcggAatccaaaactc
Fragment 6
2351-GP64_dBamH1_2-F gagttttggatTccgaccatcggcaac
2351-GP64_Xho1-R (gaaaccgtaatagacaatattaa ctcgag atagca)
tgctatctcgagttaatattgtctattacggtttc
jion fragments by PCR
DIGEST WITH NdeI+XhoI
Colony Screening
[edit]Date: 2014_8
Make miniprep, use NdeI + XhoI to find positive clone
