Team:Cambridge/Lab book/Week 7

From 2012.igem.org

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===Tuesday===
===Tuesday===
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'''[[zzz|Production of electrocompetent e.coli]]'''
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'''[[Team:Cambridge/Protocols/Electrocompetentcells|Production of electrocompetent e.coli]]'''
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'''[[Team:Cambridge/Protocols/Chemicallycompetentcells|Production of chemically competent e.coli]]'''
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===Wednesday===
===Wednesday===

Revision as of 14:51, 7 August 2012

Week: 3 4 5 6 7

Contents

Monday

PCR of Magnesium riboswitch vector fragment B and Magnesium promoter


Riboswitch and promoter gel. Lanes 2+3: Fragment B without 8 codon substitution, lanes 4+5: Fragment B with 8 codon substitution, lanes 6+7: magnesium sensitive promoter, lane 8: +ve control
  • Normal PCR settings used, annealing temperature 57 °C, elongation step 90s long.
  • Lane 5 accidentally loaded with a DNA ladder instead of loading dye.
  • Expected fragment sizes:
  • Lane 2-5: 3kbp
  • Lane 6-7: 300bp
  • After electrophoresis, found vector products had, for the most part, worked. Promoter elements were not amplified - no band of the expected size was observed.
  • Positive control also failed, although this has ceased to work for several days, potentially due to DNA degradation.

Tuesday

Production of electrocompetent e.coli


Production of chemically competent e.coli


Wednesday

Thursday

Friday