IGEM:Groningen/Notebook/iGEM 2011/2011/07/25: Difference between revisions
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<br> Plates of transformants with PBAD-cI-LVA-DT and PBAD-RBS-GFP-DT vector look good, since the self ligation plates don't have much colonies! | <br> Plates of transformants with PBAD-cI-LVA-DT and PBAD-RBS-GFP-DT vector look good, since the self ligation plates don't have much colonies! | ||
<br> 10μl plate is empty on self ligation plate control! 90μl plate has a few colonies... | <br> 10μl plate is empty on self ligation plate control! 90μl plate has a few colonies... | ||
<br> Colony PCR as usual | <br> Colony PCR as usual-> PBAD-cI-LVA-DT looks OK-> grow colony 8 and 9 overnight. | ||
<br> | <br> | ||
<br> Digestion PBAD, PhybB and vectors: cI-LVA and RBS-GFP | <br> Digestion PBAD, PhybB and vectors: cI-LVA and RBS-GFP | ||
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<br> Digestion vector (with LasR-LVA-DT): | <br> Digestion vector (with LasR-LVA-DT): | ||
<br> | <br> | ||
<br> | <br> 3μl vector | ||
<br> 1μl EcoRI | <br> 1μl EcoRI | ||
<br> 1μl XbaI | <br> 1μl XbaI | ||
<br> 3μl Fast digest buffer | <br> 3μl Fast digest buffer | ||
<br> 1μl Fast AP | <br> 1μl Fast AP | ||
<br> | <br> 21μl MQ water | ||
<br> | <br> | ||
<br> Ligation: | <br> Ligation: | ||
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<br> Transformation as usual | <br> Transformation as usual | ||
<br> | <br> | ||
<br> Plasmid prep of LasR-LVA-DT colonies 6 and 7. DNA Concentration according to the N-D1000 Nanodrop is 46.7 and 51 ng/μl <br> respectively. | |||
<br> Colony 6 plasmid prep sample was used for vector sample today | |||
<br> Colonies 6 and 7 were send for sequencing and a glycerol stock has been made. | |||
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Revision as of 07:22, 25 July 2011
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25-7-11
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