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523-pCDNA3.1-Bcl-2 - Revision history
2026-10-18T12:08:28Z
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Paul.xu at 14:46, 19 March 2012
2012-03-19T14:46:50Z
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<p><b>New page</b></p><div>Cloning of pCDNA3.1-Bcl-2 (523) <br />
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Purpose: to clone pCDNA3.1-Bcl-2 by Gene Synthesis <br />
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Date: 2012-1-18 to 2012-1-23 <br />
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Notebook: Xiaojing Wang <br />
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Page: 004163---004164 <br />
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Clone ID: 523 <br />
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Experimenters: Xiaojing Wang <br />
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Results <br> <br />
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[[Image:523 PCR 1.20.2012 S.JPG]]<br> <br />
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digest:<br> <br />
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[[Image:523 and vector digest 1.19.2012.JPG]]<br> <br />
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digest PCR product and vector with Hind111 +Xho1,run gel <br />
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the left band shows 523 insert, the right picture shows 335 vector (pCDNA3.1, 335) <br />
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gel purify, ligation, transformation <br />
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<br>Colony Screening <br />
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[[Image:523 digest 1.23.2012.JPG]]<br> <br />
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Select 5 clones to mini-prep, digestd by Hind111 and Xho1. 523-1, -2, -3, -4 may be correct. Control is 523-1. <br />
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Select 523-1 to sequence.<br> <br />
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<br> <br />
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Cloning of pCDNA3.1-Bcl-2 (523) <br />
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Purpose: to clone pCDNA3.1-Bcl-2 by Gene Synthesis <br />
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Date: 2012-3-13 to 2012-3-16 <br />
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Notebook: <br> <br />
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Page: <br> <br />
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Clone ID: 523 <br />
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Experimenters: yingying lu<br> <br />
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Results <br> <br />
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[[Image:523pPCR 3-12-2012.JPG]]<br> <br />
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[[Image:523pPCR 3-13-2012.JPG]]<br> <br />
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PCR:PCR using pDONR-Bcl-2, 1019 as template <br />
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oligos <br />
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Bcl-2-F C6 tgatcaAAGCTTGAATTCGCCGCCACCATG 10 uM<br>Bcl-2-2 CCGGTTATCGTACCCTGTTCTCCCAGCGTGCGCCATGGTGGCGGCGAATTCGTA 1 uM<br>Bcl-2-3 D8 GAACAGGGTACGATAACCGGGAGATAGTGATGAAGTACATCCATTATAAGCTGT 1 uM <br />
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<br>and <br>Bcl-2-20 CACCCAGGGTGATGCAAGCTCCCACCAGGGCCAAACTGAGCAGAGTCTTCAGAG 1 uM<br>Bcl-2-21 CTTGCATCACCCTGGGTGCCTATCTGGGCCACAAGTGAGAATTCGCTCTCGAGG 1 uM<br>Bcl-2-R C7 tactcCTCGAGGAATTCTCACTTGTGGCCCAG 10 uM<br> <br />
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[[Image:523 clone 3-14-2012.JPG]]<br> <br />
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{| cellspacing="1" cellpadding="1" border="1" style="width: 396px; height: 115px;"<br />
|-<br />
| <br> <br />
| band1<br> <br />
| band 2<br> <br />
| band 3<br> <br />
| band 4<br><br />
|-<br />
| <br> <br />
| PCR product 3.12<br> <br />
| 335pCDNA3.1<br> <br />
| PCR product 3.13<br> <br />
| 335pCDNA3.1<br><br />
|-<br />
| digest enzymes<br> <br />
| colspan="2" | Hind111 + Xho1<br> <br />
| colspan="2" | Hind111 + Xho1<br><br />
|}<br />
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[[Image:523 digest detect 3-16-2012.JPG]]<br> <br />
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clonies digest detect: Hind111 + Xho1<br />
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[[Image:523_digest_detect_3-19-2012.JPG]]<br />
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digest detect again:EcoR1<br />
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--[[User:Paul.xu|Paul.xu]] 14:56, 16 March 2012 (EST) <br />
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[[Category:Cloning]]</div>
Paul.xu