IGEM:University of Groningen/2011/Notebook/Count coli/2011/08/10

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Count coli Project description
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Wed 10 August 2011

To Do List

  • Transformation---2nd batch (ON ligation mixture)
  • Screening transforman ---Growth ON culture:
    • 12 variant autoinducing construts
    • RFP internal control --> PCR colonies screening
    • construct 21.12 (pSB1K3 revDT+P(LasR)+LasR-TAG)

Implementation RBS-RFP PCR colonies screening --> pick 9 single clones
15.5 ul MQ
2 ul buffer
1.2 ul MgCl2
0.4 ul dNTPs
0.4 ul Forward
0.4 ul Reverse
0.1 ul Taq DNA pol
DNA sample

Growth ON culture for plasmid isolation from
7 plates 22.xx : 2 clones each
1 plate 21.12 : 2 clones

Results None of growing clones have RBS-RFP insert --> self-ligation/incomplete vector digestion


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