1251 pET28b-KOD-sso7d: Difference between revisions
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Latest revision as of 11:46, 19 July 2012
Cloning of 1251 pET28b-KOD-sso7d
[edit]Purpose: Cloning of 1251 pET28b-KOD-sso7d
Date: July 12, 2012
Notebook:
Page:
Clone ID: 1251
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: July 12, 2012
Digested 1250 pET28b-KOD-2 with Nde1 + Kpn1 --> insert (lane 1, bottom band)
Digested 368 pET30b-sso7d-KX with Nde1 + Kpn1 -->vector (lane 2, top band)
Date: July 17, 2012
MiniPrep digestion was strange, digest insert and vector again. Only two bands this time, instead of three. Do not know why there were three bands before, maybe used the wrong enzymes. Ligate together
Digested 1250 pET28b-KOD-2 with Nde1 + Kpn1 --> insert (lane 1, bottom band)
Digested 368 pET30b-sso7d-KX with Nde1 + Kpn1 -->vector (lane 2, top band)
Cut out band from gel, purify, ligated for 1 hour and transform with Top10 cells and plated on Kan plates to grow over night
Colony Screening
[edit]Date: July 16, 2012
Digested MiniPrep with Nde1 + Kpn1
July 17, 2012
Digested MiniPrep again with Nde1 + Kpn1 a little bit longer. Maybe there was something wrong with the digested before, should ligate together and transform again
Date: July 19, 2012
Digested MiniPrep again with Nde1 + Kpn1 for 1 hr 15 minutes
Result intepretation/Summary
[edit]Conclusion
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