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2350 pFastbac-GP64 - Revision history
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Excellgen at 15:58, 29 October 2014
2014-10-29T15:58:49Z
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<p><b>New page</b></p><div>== '''Cloning of 2350 pFastbac-GP64''' ==<br />
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'''Purpose:''' Cloning of2350 pFastbac-GP64'''==''' <br />
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'''Date: '''2014-9---- <br />
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'''Notebook:0''' <br />
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'''Page:''' <br />
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'''Clone ID:2350''' <br />
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'''Experimenters:'''&nbsp; shanyi <br />
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== '''Results''' ==<br />
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=== CLONING ===<br />
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PCR from 2351 pEC2-GP64 <br />
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Fragment 1 <br />
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2350-GP64-Xho1-F tacgac ctcgag ATGGTAAGCGCTATTGTTTTATATG <br />
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2350-GP64-R1 (gaaaccgtaatagacaatattaa gtac) gtacttaatattgtctattacggtttc <br />
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<br> Fragment 2 <br />
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2350-GP64-Xho1-F tacgac ctcgag ATGGTAAGCGCTATTGTTTTATATG <br />
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2350-GP64-R2 (gaaaccgtaatagacaatattaa) ttaatattgtctattacggtttc <br />
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[[Image:2350 1917 2240pcr.JPG]]<br> <br />
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purify two fragments in one tube, heat to 95 oC, cool slowly to RT then digest with Xho1 (without Kpn1) <br />
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ligate to Xho1 + Kpn1 digested vector <br />
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[[Image:Digest for 2350 2367 2368.JPG]]<br> <br />
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=== Colony Screening ===<br />
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Date: 2014_8<br> <br />
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Make miniprep, use Xho1 + Kpn1 to find positive clone<br> <br />
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[[Image:2240 2350 2367 2368diget.JPG]]<br> <br> <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>
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