Melb:Plan:GFP Fluorescent reporter

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Revision as of 07:41, 11 August 2007 by Patricia (Talk | contribs)

LacZ reporter

  1. Ligate LacZ coding region to gentamycin RBS part. (Done?)(1 day)
  2. Transform and select for appropriate construct using Kan/Gen plate. (Done?)(1 day + overnight)
  3. Liquid culture, miniprep and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (overnight + 1 day)
  4. Make glycerol stock of RBS-LacZ containing cells. (same day)
  1. Ligate the RBS-LacZ construct into double terminator plasmid. (same day)
  2. Transform ligation and select for construct using Gen/Amp plate. (1 day + overnight)
  3. Liquid culture, miniprep and confirm using EcoRI/Pst digest (overnight + 1 day)
  4. prepare glycerol stock (same day)


This construct can then be ligated to appropriate promoters for use as a test harness