1277 pET30b-RNaseIII
Cloning of 1277 pET30b-RNaseIII
[edit]Purpose: Cloning of 1277 pET30b-RNaseIII
Date: July 30, 2012
Notebook:
Page:
Clone ID: 1277
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: July 30, 2012
See lane 4
PCR using genomic DNA from K-12 substr. MG1655
RnaseIII-Nde1-F AGCTCTCATatgaaccccatcgtaattaatc
RnaseIII-Xho1-R gtctcgctcgagttccagctccagttttttcaac
clone into Nde1 + Xho1 digested 1254 pTXB1-PT-ELK
Expected size 1311 bp
Cut out band and save in box
Date: August 1, 2012
136
Digested insert (lane 4) with RE Nde1 + Xho1, vector is not good, digest again tomorrow, join with lane 4, purify and ligate together
Colony Screening
[edit]Date: August 6, 2012
Digested MiniPrep with RE Xho1 and Nde1, expected size 1311 bp
136
All negative for inserts
PCR
DIGEST V15pET30b and 1277pcr produts with Nde1 + Xho1
SCREENING
Result intepretation/Summary
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Conclusion
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