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359-pET30-pMBP - Revision history
2026-10-18T05:43:31Z
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Luyingying at 20:31, 10 November 2011
2011-11-10T20:31:23Z
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<p><b>New page</b></p><div>Cloning of 359-pET30-pMBP<br> <br />
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Purpose: to clone pET30-pMBP by PCR <br />
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Date: 2011-10-4---- <br />
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Notebook:Lu yingying <br />
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Page: <br />
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Clone ID:359 <br />
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Experimenters: lu yingying<br>Results<br> <br />
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[[Image:359-pMBP-PCR-product10-4-2011.JPG]]<br> <br />
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PCR product of pMBP<br> <br />
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template: 424-pDEST-periHisMBP<br> <br />
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primer:periMBP-F + 10% periMBP-R1 + periMBP-R2<br> <br />
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25 cycles <br />
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[[Image:359pMBP+PET30b digest 10-5-2011.JPG]]<br> <br />
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digest result:NdeI + NcoI<br> <br />
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band 1 shows pMBP digest result, band 2 shows PET30b digest result<br> <br />
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[[Image:344,359,391+393digest detect10-7-2011.JPG|519x241px|344,359,391+393digest detect10-7-2011.JPG]] <br />
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359,1-6 bands show digest result: no insert <br />
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Cloning of 359-pET30-pMBP <br />
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Purpose: to clone pET30-pMBP by PCR <br />
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Date: 2011-10-17---- <br />
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Notebook:Lu yingying <br />
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Page: <br />
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Clone ID:359 <br />
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Experimenters: lu yingying<br>Results<br> <br />
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[[Image:359-PMBP10-17-2011.JPG]]<br> <br />
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PCR product of pMBP <br />
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template: 424-pDEST-periHisMBP <br />
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primer:periMBP-F + 10% periMBP-R1 + periMBP-R2 <br />
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25 cycles <br> <br />
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[[Image:359-PMBP digest10-17-2011.JPG]]<br> <br />
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digest result:NdeI + NcoI<br> <br />
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[[Image:359-PET30b digest-10-17-2011.JPG]]<br> <br />
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digest result:NdeI + NcoI<br> <br />
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ligation RT overnight <br />
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Cloning of 359-pET30-pMBP <br />
<br />
Purpose: to clone pET30-pMBP by PCR <br />
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Date: 2011-10-19---- <br />
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Notebook:Lu yingying <br />
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Page: <br />
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Clone ID:359 <br />
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Experimenters: lu yingying<br>Results<br> <br />
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[[Image:359-PMBP PCR product10-19-2011.JPG]]<br> <br />
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[[Image:359-vector+pMBP digest10-20-2011.JPG]]<br> <br />
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digest result:NdeI + NcoI NEB enzyme for 2 hours<br> <br />
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Date: 2011-10-25---- <br />
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Notebook:Lu yingying <br />
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Page: <br />
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Clone ID:359 <br />
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Experimenters: lu yingying<br>Results<br> <br />
<br />
[[Image:359-PMBP PCR product10-19-2011.JPG]]<br> <br />
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PCR product of pMBP <br />
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template: 424-pDEST-periHisMBP <br />
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primer:periMBP-F + 10% periMBP-R1 + periMBP-R2 <br />
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25 cycles <br> <br />
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10-19 PCR product 10ul left<br> <br />
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[[Image:359 subclone 10-26-2011.JPG]]<br> <br />
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digest result:NdeI + NcoI&nbsp; <br> <br />
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[[Image:414+470+375+359 digest detect10-27-2011.JPG]]<br> <br />
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359 digest detect: NdeI + NcoI<br> <br />
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Date: 2011-11-4----2011-11-7 <br />
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Notebook:Lu yingying <br />
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Page:003966----003971 <br />
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Clone ID:359 <br />
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Experimenters: lu yingying<br>Results<br> <br />
<br />
[[Image:359-PMBP PCR product10-19-2011.JPG]]<br> <br />
<br />
PCR product of pMBP <br />
<br />
template: 424-pDEST-periHisMBP <br />
<br />
primer:periMBP-F + 10% periMBP-R1 + periMBP-R2 <br />
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25 cycles <br />
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10-19 PCR product 10ul left<br> <br />
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[[Image:375+359_clone_11-4-2011.JPG]]<br><br />
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digest result:NdeI + NcoI <br><br />
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[[Image:359+375_digest_detect11-7-2011.JPG]]<br><br />
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5 clonies digest detect:NdeI + NcoI ,need to sequence to detect<br />
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[[Category:Cloning]]</div>
Luyingying