Team:Cambridge/Lab book/Week 8

From 2012.igem.org

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===Tuesday (14/08/12)===
===Tuesday (14/08/12)===
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[[Team:Cambridge/Protocols/Gibsonassembly|Gibson assembly of positive control]]
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*Protocol changed slightly: 0.5 μl of each DNA fragment solution mixed in with master mix to make up 4 μl total volume (
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[[Team:Cambridge/Protocols/TransformationofE.coli|Transformation of e.coli with positive control DNA]]
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----
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*Chemically competent e.coli cells transformed with plasmid DNA produced by Gibson assembly step.
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*Transformants plated out on 50 μg/ml kanomycin plates. Incubated overnight at 37 °C.
===Wednesday (15/08/12)===
===Wednesday (15/08/12)===

Revision as of 10:12, 15 August 2012

Week: 3 4 5 6 7 8 9

Contents

Monday (13/08/12)

Tuesday (14/08/12)

Gibson assembly of positive control

  • Protocol changed slightly: 0.5 μl of each DNA fragment solution mixed in with master mix to make up 4 μl total volume (

Transformation of e.coli with positive control DNA


  • Chemically competent e.coli cells transformed with plasmid DNA produced by Gibson assembly step.
  • Transformants plated out on 50 μg/ml kanomycin plates. Incubated overnight at 37 °C.

Wednesday (15/08/12)

PCR of positive control fragments


  • Standard PCR settings used

Extraction of positive control DNA


  • Positive control DNA from gel excised and purified.
  • Additional elution steps used to concentrate resultant solution.
  • Verified with nanodropper - final DNA concentrations:

Thursday (16/08/12)

Friday (17/08/12)