1141 p8H-DNA-Pol-1
Cloning of 1141 p8H-DNA-Pol-1
[edit]Purpose: Cloning of 1141 p8H-DNA-Pol-1
Date: July 23, 2012
Notebook:
Page:
Clone ID: 1141
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: July 27, 2012
See lane 5, expected size 2788 bp
PCR using MG1665 genomic DNA
DNA-Pol1-Nde1-F
catgtaCATATGgttcagatcccccaaaatc
DNA-Pol1-Sal1-R
(cgaaaactgggatcaggcgcac GTC GAC tca cgat)
atcgtgaGTCGACGCGGCCGCCgtgcgcctgatcccagttttcg
Digest into vector 1135 with RE Nde1 and Sal1
See lane 10 (insert) and lane 11 (vector)
Cut out bands and ligate together and transform
Date: August 2, 2012
Screening came back bad, started all over again
See lane 4, expected size 2788 bp
PCR using MG1665 genomic DNA
DNA-Pol1-Nde1-F
catgtaCATATGgttcagatcccccaaaatc
DNA-Pol1-Sal1-R
(cgaaaactgggatcaggcgcac GTC GAC tca cgat)
atcgtgaGTCGACGCGGCCGCCgtgcgcctgatcccagttttcg
1
Ligate together, transform
Digest into vector 1135 with RE Nde1 and Sal1
Colony Screening
[edit]Date: July 27, 2012
Digested MiniPrep with RE Nde1 and Sal1, expected size 2788 bp
Samples 2, 5 and 6 may be correct, submitted sample 5 for sequencing
Date: August 6, 2012
1
Digested MiniPrep with RE Nde1 and Sal1, expected size 2788 bp
No inserts
Date: Nov30, 2012
Digest vector 1135 and 1141pcr products with Nde1 and Sal1
Digested MiniPrep with RE Nde1 and Sal1, expected size 2788 bp
Result intepretation/Summary
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Conclusion
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