Team:UGent/Labjournal
From 2013.igem.org
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<li><b>Restriction</b>of ccdB operon, pSB3T5 and pSB4A5 using PstI-HF and XbaI (gel purify RD-fragments using Qiagen Qiaquick gel extraction kit (expected fragment of 2157 bp for T7-ccdB, 3229 bp for pSB3T5, 3372 bp for pSB4A5 and 3999 bp for pSB6A1): concentrations deduced from gel</li> | <li><b>Restriction</b>of ccdB operon, pSB3T5 and pSB4A5 using PstI-HF and XbaI (gel purify RD-fragments using Qiagen Qiaquick gel extraction kit (expected fragment of 2157 bp for T7-ccdB, 3229 bp for pSB3T5, 3372 bp for pSB4A5 and 3999 bp for pSB6A1): concentrations deduced from gel</li> | ||
<li><b>Ligation</b> of pSB3T5 and T7ccdB (from p5SpFRT-T7ccdB), pSB4A5 and T7ccdB (from p5SpFRT-T7ccdB) and pSB4A5 and T7ccdB (from p20SpFRT-T7ccdB) with T4 DNA ligase at 16°C overnight</li> | <li><b>Ligation</b> of pSB3T5 and T7ccdB (from p5SpFRT-T7ccdB), pSB4A5 and T7ccdB (from p5SpFRT-T7ccdB) and pSB4A5 and T7ccdB (from p20SpFRT-T7ccdB) with T4 DNA ligase at 16°C overnight</li> | ||
- | <li><b>Transformation</b> of ligation mixtures in DH5a using heat shock</li> | + | <li><b>Transformation</b> of ligation mixtures in DH5a using heat shock |
+ | <br>-> no colonies</li> | ||
</ul> | </ul> | ||
<br> | <br> |
Revision as of 10:56, 5 September 2013
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