Melb:Plan:GFP Fluorescent reporter
From 2007.igem.org
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==LacZ reporter== | ==LacZ reporter== | ||
| - | #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 | + | #Liquid culture, miniprep and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (overnight + 1 day) |
| - | #Make glycerol stock of RBS-LacZ containing cells. ( | + | #Make glycerol stock of RBS-LacZ containing cells. (same day) |
| - | #Ligate the RBS-LacZ construct into double terminator plasmid. ( | + | #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
- Ligate LacZ coding region to gentamycin RBS part. (Done?)(1 day)
- Transform and select for appropriate construct using Kan/Gen plate. (Done?)(1 day + overnight)
- Liquid culture, miniprep and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (overnight + 1 day)
- Make glycerol stock of RBS-LacZ containing cells. (same day)
- Ligate the RBS-LacZ construct into double terminator plasmid. (same day)
- 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
