2267 pT7A3-Tth-PPase
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Cloning of 2267 pT7A3-Tth-PPase
[edit]Purpose: Cloning of2267 pT7A3-Tth-PPase==
Date: 2014-9----
Notebook:0
Page:
Clone ID:2267
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from Thermus thermophilus HB27 genomic DNA
Fragment 1
Tth-PPase-Nde1-F
atcgac catatggcgaacctgaagagccttc
2267-Tth-PPas-R1 cttggcctcCagggccttg
Fragment 2
2267-Tth-PPas-F1 caaggccctGgaggccaag
Tth-PPase-Xho1-R
gtgctactcgaggcccttgtagcgggcgatgcag
(ctgcatcgcccgctacaagggc ctcgag tagcac)
Join 2 fragments
Tth-PPase-Nde1-F
atcgac catatggcgaacctgaagagccttc
Tth-PPase-Xho1-R
gtgctactcgaggcccttgtagcgggcgatgcag
(ctgcatcgcccgctacaagggc ctcgag tagcac
digest with Nde1 + Xho1
Colony Screening
[edit]Date: 2014_8
Make miniprep, use Nde1 + Xho1to find positive clone
