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1714 pEC-iMMLV - Revision history
2026-10-18T07:06:29Z
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Paul.xu at 18:07, 10 April 2013
2013-04-10T18:07:42Z
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<p><b>New page</b></p><div>== '''Cloning of 1714 pEC-iMMLV''' ==<br />
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'''Purpose:''' Cloning of1714 pEC-iMMLV <br />
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'''Date: '''2013-4- <br />
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'''Notebook:''' <br />
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'''Page:''' <br />
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'''Clone ID:1714''' <br />
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'''Experimenters:''' Shanyi <br />
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== '''Results''' ==<br />
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=== cloning ===<br />
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Date:2013-3-<br> <br />
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PCR use 1713 as template and primers <br> <br />
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MMLV-EcoR1-Nde1-F + MMLV-E69K-R <br> <br />
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MMLV-E69K-F + MMLV-E302R-R <br> <br />
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MMLV-E302R-F + MMLV-N454K-R <br> <br />
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MMLV-N454K-F + MMLV-D524N-R <br> <br />
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MMLV-D524N-F + MMLV-Not1-Xho1-R <br />
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[[Image:1705 1714pcr.JPG]]<br> <br />
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join 5 fragments by pcr using<br> <br />
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MMLV-EcoR1-Nde1-F atcgac gaattc CAT ATG ACCCTAAATATAGAAGATGAGC <br />
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MMLV-Not1-Xho1-R (CTCCAGACACCTCTACCCTCCTC cat cac cat cac cat cac taa GCGGCCGC CTCGAG cgatca) tgatcgCTCGAGGCGGCCGCttagtgatggtgatggtgatgGAGGAGGGTAGAGGTGTCTGGAG <br />
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[[Image:1714pcr.JPG]]<br> <br />
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digest with&nbsp; EcoR1 + Xho1 <br />
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[[Image:Digest for 1714first five mutation.JPG]]<br> <br />
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{| cellspacing="1" cellpadding="1" border="1"<br />
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=== Colony Screening ===<br />
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Date: 2013 <br />
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DIGEST WITH BamHI<br> <br />
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[[Image:1714_1732digest.JPG ]]<br />
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== '''Result intepretation/Summary''' ==<br />
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*<br><br />
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== '''Conclusion''' ==<br />
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[[Category:Cloning]]</div>
Paul.xu