Team:Groningen/Notebook/9 July 2009
From 2009.igem.org
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Revision as of 12:10, 17 August 2009
Wet
GVP Cluster
Inoculation of TY-medium for Glycerol Stocks and Plasmid isolation
- BBa_J23109 (labelled as no.1) vector BBa_J61002 in E.coli TOP10
- BBa_J23100 (labelled as no.2) vector BBa_J61002 in E.coli TOP10 (red colony)
- BBa_J23106 (labelled as no.3) vector BBa_J61002 in E.coli TOP10 (red colony)
- The three plates showing new colonies were used to inoculate 5mL TY-medium with amp. (final conc. 100mg/ul).
- The tubes were put in a waterbath at 37C (200 rpm) and grown o.n.
Transporters
Purification of HmtA (PA Q9I147 6xHis Tag)
- SDS-gel electrophosis
- Make a 12% SDS-gel
- Fill gel with samples Wash 1-2 20mM imidazole, Wash1-2 40mM imidazole, Elute1-3 and ladders
- Run 2h at 100V
- Stain the gel in Coomassie blue (2h)
- Destain with demiwater (2x, 10 min microwave)
Vectors
Measure concentration with Nanodrop
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