PLenti-LSS-mKate2: Difference between revisions
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Latest revision as of 16:09, 1 September 2011
SubCloning of LSS-mkate2 to pLentiplus
[edit]Purpose: mutagenesis of LSS-mkate2 using pet28-mkate2 as template and subclone to plentiplus.
Date: 2011-8-11
Notebook: Haiyan
Page: 003021
Clone ID: 218
Experimenters: Haiyan
Results
[edit]using DpnI and Turbo DNA Polymerase, pet28-mkate2 as template, transform to top 10, kana
5ul and 15 ul pcr product both got the colonies. pick up 6 colonies to do miniprep
lane 4-10, pet28-lss-mkate 2 bamhi+xhoi cut
lane 1-3, pet 28-yfp bamhi+xhoi cut, throw away clone 1,2, just keep 3, freeze bacteria.
Subcloning
[edit]Date: 2011-6-21
PCR 1A) mKate1+LSS-mKateR primers
PCR 1B) LSS-mKateF + mKate20 primers
Purified and mixed 2 fragments, did PCR2 with mKate1+mKate20 primers
Digested DNA and pLentiplus vector w/ BamH1 and Xho1
Ligation and STBL3 Transformation
Colony Screening
[edit]Date:2011-8-30
Miniprep Plenti-Lss-mKate2, and then use BamHI + XhoI to digest:
Lane 7-9 show the result.
Result intepretation/Summary
[edit]Conclusion
[edit]Paul.xu 08:23, 15 June 2011 (EST)