1297 pET-6H-VPMLK-TAT-NLS
Cloning of 1297 pET-6H-VPMLK-TAT-NLS
[edit]Purpose: Cloning of 1297 pET-6H-VPMLK-TAT-NLS
Date: August 13, 2012
Notebook:
Page:
Clone ID: 1297
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: August 13, 2012\
Gene synthesis
Purify PCR product, see lane 2, (113 bp, need to add 10% isopropanol becasue the fragment is very small).
Digest PCR product with RE Nco1 + Nde1.
Digest vector 1128 pDB.His.PfuMBP-(tev) with Nco1 + Nde1.
Ligate together overnight and transform
Date: August 14, 2012
Insert disappeared after digestion, need to reamplify, see lane 1
Date: August 15, 2012
Digest PCR product with RE Nco1 + Nde1 (lane 1)
Digest vector pDB.His.MBP, V14 with Nco1 + Nde1 (lane 2)
Ligate together and transform
Colony Screening
[edit]Date: August 20, 2012
Digested MiniPrep with RE Nco1 and Nde1
Faint band at expected size (~100 bp) maybe send to sequencing
Result intepretation/Summary
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Conclusion
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