Brown: Team Resistance/22 October 2008

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(Difference between revisions)
(New page: {{BNBK}} ==22 October 2008== ====Ligation==== pVJ4 lysis cassette and S105 lysis cassette into standard biobrick vector plasmid pSB1AK3 at EcoRI and SpeI *15 ul mixes: *12.37ul diH20 *1...)
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#control (no ara*)
#control (no ara*)
#test (480ul ara*)
#test (480ul ara*)
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Because the cells grew during the experiment into stationary phase, the lysis seems to take much longer than usual.

Revision as of 17:38, 22 October 2008


Toxipop lab notebook.png

Contents

22 October 2008

Ligation

pVJ4 lysis cassette and S105 lysis cassette into standard biobrick vector plasmid pSB1AK3 at EcoRI and SpeI

  • 15 ul mixes:
  • 12.37ul diH20
  • 1.5ul T4 10x buffer
  • 2.08ul pVJ4 OR

2.62ul S105

  • 0.53uL vector
  • 0.5ul T2 ligase

Transformation

  • Used 50ul competent cells (invitrogen)
  • skipped the SOC step (immediately plated) since SOC step not necessary for amp resistant plasmid
  • Put in incubator at 10:30am

Conductance test

Measuring conductance change and optical density change in parallel

Set up two cultures of the pVJ4 plasmid

  1. control (no ara*)
  2. test (480ul ara*)

Because the cells grew during the experiment into stationary phase, the lysis seems to take much longer than usual.

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