Team:Groningen/Notebook/15 July 2009
From 2009.igem.org
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m (→GVP Cluster) |
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* 1μL SpeI fast digest enzyme | * 1μL SpeI fast digest enzyme | ||
* 1μL PstI fast digest enzyme | * 1μL PstI fast digest enzyme | ||
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+ | The cups were incubated in a heat block at 37°C for 30 min. followed by adding 4μL 6x loading buffer for gel electroforesis. | ||
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'''Gel electroforesis''' | '''Gel electroforesis''' |
Revision as of 07:46, 15 July 2009
Wet
GVP Cluster
Restriction for Assembly
The vector (BBa_J61035) containing GVP cluster was cut with PstI and XbaI to create correct ends for insert in promotor vector (BBa_J61002).
- 10μL MQ
- 6μL plasmid in MQ (2μg)
- 2μL Fast digest buffer
- 1μL PstI fast digest enzyme
- 1μL XbaI fast digest enzyme
The vector (BBa_J61002) containing promotor BBa_J23100 was cut with SpeI and PstI.
- 4μL MQ
- 10μL plasmid in MQ
- 2μL Fast digest buffer
- 1μL SpeI fast digest enzyme
- 1μL PstI fast digest enzyme
The cups were incubated in a heat block at 37°C for 30 min. followed by adding 4μL 6x loading buffer for gel electroforesis.
Gel electroforesis
24μL of each sample was loaded on a 1% agarose gel with EtBr and a 1kb ladder was used (see picture).
- → From left to right: 1kb Marker,
Transporters
Dry
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