1117 p8H-Klenow-Exo-
Cloning of 1117 p8H-Klenow-Exo-
[edit]Purpose: Cloning of 1117 p8H-Klenow-Exo-
Date: July 23, 2012
Notebook:
Page:
Clone ID: 1117
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: July 23, 2012
PCR using MG1665 genomic DNA
KlenowExo-_Nde1-F
cagtcg catatg ATTGAACGTAACGGTGTGAAG
KlenowExo-_Xho1-R
(GAAAACTGGGATCAGGCGCAC CTCGAG ctgtca)
tgacagCTCGAGGTGCGCCTGATCCCAGTTTTC
Expected size 1168 bp
Cut out bands in lanes 1 and 2, purify and digest with RE Nde1 and Xho1 with vector p8H
See lanes 2 and 3, cut out both bands and ligate together and transform
Date: August 2, 2012
Sequencing came back bad, start over again
PCR using MG1665 genomic DNA
KlenowExo-_Nde1-F
cagtcg catatg ATTGAACGTAACGGTGTGAAG
KlenowExo-_Xho1-R
(GAAAACTGGGATCAGGCGCAC CTCGAG ctgtca)
tgacagCTCGAGGTGCGCCTGATCCCAGTTTTC
Expected size 1168 bp
Cut out bands in lanes 1, purify and digest with RE Nde1 and Xho1 with vector p8H
1
See lanes 1 and 2, cut out both bands and ligate together and transform
Colony Screening
[edit]Date: July 27, 2012
Digested MiniPrep with RE Nde1 and Xho1. See lanes 1-6, expected size ~1160 bp. Samples 2 and 4 may be correc, send sample 2 to sequencing.
Date: August 6, 2012
Digested MiniPrep with RE Nde1 and Xho1. See lanes 1-6, expected size ~1160 bp
Lanes 1, 2, 3, 4 may be correct
Send sample 2 to sequencing
Purpose: Cloning of 1117 p8H-Klenow-Exo-
Date: Dec, 2012
Notebook:
Page:
Clone ID: 1117
Experimenters: Shanyi
PCR
DIGEST 1422 and 1277 pcr products with Nde1 + Xho1
SCREENING
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Result intepretation/Summary
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Conclusion
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