IGEM:Groningen/Notebook/iGEM 2011/2011/06/08: Difference between revisions
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== | ==08-06-11== | ||
<br> | |||
<br> Biobricks were checked (by sequencing) | |||
<br> | |||
<br> Transform: Digestion analysis: Sequencing: | |||
<br> PlasB: OK OK | |||
<br> RFP: OK OK | |||
<br> cILVA: OK OK | |||
<br> DT: OK OK | |||
<br> rev DT: OK OK | |||
<br> PBADaraC: OK OK | |||
<br> PcI: - - | |||
<br> GFP-LVA: - - | |||
<br> RBS-GFP-DT: OK OK | |||
<br> | |||
<br> Plan: Digestion of RBS-GFP-DT with EcoRI and XbaI | |||
<br> PlasB and PBADaraC with EcoRI and SpeI | |||
<br> isolate both of gel and use also normal PBADaraC sample for ligation: | |||
<br> | |||
<br> Digestion vector: | |||
<br> 2μl FD buffer | |||
<br> 0.5μl EcoRI | |||
<br> 0.5μl XbaI | |||
<br> 7μl MQ water | |||
<br> 10μl mix+ 10μl sample | |||
<br> | |||
<br> Digestion mix insert: | |||
<br> 14μl FD buffer | |||
<br> 3.5μl EcoRI | |||
<br> 3.5μl SpeI | |||
<br> 49μl MQ water | |||
<br> | |||
<br> Digest for 45 min at 37 degrees | |||
<br> | |||
<br> Clean up of the DNA with the PCR purification kit | |||
<br> | |||
<br> Ligation: | |||
<br> 7μl insert | |||
<br> 3μl plasmid | |||
<br> 2μl T4 DNA ligase buffer | |||
<br> 1μl T4 DNA ligase | |||
<br> 7μl MQ water | |||
<br> | |||
<br> Ligate for 2hours at room temperature and afterwards,ligate overnight | |||
<br> | |||
Revision as of 02:12, 21 September 2011
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08-06-11
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