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PLenti-LSS-mKate2 - Revision history
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Ruili.su at 20:09, 1 September 2011
2011-09-01T20:09:38Z
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<p><b>New page</b></p><div>== '''SubCloning of LSS-mkate2 to pLentiplus''' ==<br />
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'''Purpose:''' mutagenesis of LSS-mkate2 using pet28-mkate2 as template and subclone to plentiplus. <br />
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'''Date: '''2011-8-11 <br />
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'''Notebook:''' Haiyan <br />
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'''Page: 003021 ''' <br />
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'''Clone ID:''' 218 <br />
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'''Experimenters:''' Haiyan <br />
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== '''Results''' ==<br />
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using DpnI and Turbo DNA Polymerase, pet28-mkate2 as template, transform to top 10, kana <br> 5ul and 15 ul pcr product both got the colonies. pick up 6 colonies to do miniprep <br> [[Image:IMG 1272.JPG]] <br> lane 4-10, pet28-lss-mkate 2 bamhi+xhoi cut <br> lane 1-3, pet 28-yfp bamhi+xhoi cut, throw away clone 1,2, just keep 3, freeze bacteria. <br> <br />
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=== Subcloning ===<br />
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Date: 2011-6-21 <br />
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PCR 1A) mKate1+LSS-mKateR primers <br />
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PCR 1B) LSS-mKateF +&nbsp;mKate20 primers <br />
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Purified and mixed 2 fragments, did PCR2 with mKate1+mKate20 primers <br />
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Digested DNA and pLentiplus vector w/ BamH1 and Xho1 <br />
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Ligation and STBL3 Transformation <br />
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=== Colony Screening ===<br />
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Date:2011-8-30<br />
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[[Image:PAAV-Lss-m Kate2-08-31-2011.JPG]]<br />
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Miniprep Plenti-Lss-mKate2, and then use BamHI + XhoI to digest:<br />
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Lane 7-9 show the result. <br />
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== '''Result intepretation/Summary''' ==<br />
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== '''Conclusion''' ==<br />
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=== ===<br />
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[[User:Paul.xu|Paul.xu]] 08:23, 15 June 2011 (EST)<br> <br />
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[[Category:Cloning]]</div>
Ruili.su