2229 pT7A3E-phoA
Jump to navigation
Jump to search
Cloning of 2229 pT7A3E-phoA
[edit]Purpose: Cloning of2229 pT7A3E-phoA==
Date: 2015-1----
Notebook:0
Page:
Clone ID:2229
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR from Thermus thermophilus HB27 genomic DNA
1. Tth-phoA-Nde1-F tcgaacatatgaagcgaagggacatcctg
Tth-phoA-dXho1-R ctcggcgtcctcCaggtccacAcccttcatg
2. Tth-phoA-dXho1-F catgaagggTgtggacctGgaggacgccgag
Tth-phoA-Xho1-R (caccccgaggacgtctgggccctcgagtatact) agtatactcgagggcccagacgtcctcggggtg
3. Gel purify and then join two fragments Tth-phoA-Nde1-F tcgaacatatgaagcgaagggacatcctg Tth-phoA-Xho1-R (caccccgaggacgtctgggccctcgagtatact) agtatactcgagggcccagacgtcctcggggtg
digest with Nde1 + EcoR1, clone to1982
Colony Screening
[edit]Date: 2015_1_
Make miniprep, use Nde1 + Nco1 to find positive clone
