Melb:Plan:GFP Fluorescent reporter

From 2007.igem.org

(Difference between revisions)
(LacZ reporter)
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==LacZ reporter==
==LacZ reporter==
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#Ligate LacZ coding region to gentamycin RBS part. (Done?)
+
#Ligate LacZ coding region to gentamycin RBS part. (Done?)(1 day)
-
#Transform and select for appropriate construct using Kan/Gen plate. (Done?)
+
#Transform and select for appropriate construct using Kan/Gen plate. (Done?)(1 day + overnight)
-
#Liquid culture and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously   (EcoRI and XbaI)   (Monday 13/08/07)
+
#Liquid culture, miniprep and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (overnight + 1 day)
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#Make glycerol stock of RBS-LacZ containing cells. (Monday 13/08/07)
+
#Make glycerol stock of RBS-LacZ containing cells. (same day)
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#Ligate the RBS-LacZ construct into double terminator plasmid. (Monday 13/08/07)
+
#Ligate the RBS-LacZ construct into double terminator plasmid. (same day)
-
#Transform ligation and select for construct  
+
#Transform ligation and select for construct using Gen/Amp plate. (1 day + overnight)
-
#
+
#Liquid culture, miniprep and confirm using EcoRI/Pst digest (overnight + 1 day)
 +
#prepare glycerol stock (same day)
 +
 
 +
 
 +
This construct can then be ligated to appropriate promoters for use as a test harness

Revision as of 07:41, 11 August 2007

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