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Team:Groningen/Notebook/13 July 2009
From 2009.igem.org
(Difference between revisions)
m (New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== '''Gel Purification of GVP''' A standard kit for PCR-product purification was used for gel purification * ..mg of gel ...) |
(→GVP Cluster) |
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===GVP Cluster=== | ===GVP Cluster=== | ||
| + | '''Restriction for Assembly''' | ||
| + | |||
| + | The vector containing GVP cluster was cut with EcoRI and XbaI to create correct ends. | ||
| + | |||
| + | * 6μL MQ | ||
| + | * 10μL plasmid in MQ | ||
| + | * 2μL Fast digest buffer | ||
| + | * 1μL EcoRI fast digest enzyme | ||
| + | * 1μL XbaI fast digest enzyme | ||
| + | |||
| + | The vectors containing promotors [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106] were cut with EcoRI and SpeI. | ||
| + | |||
| + | * 0μL MQ | ||
| + | * 16μL plasmid in MQ ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106]) | ||
| + | * 2μL Fast digest buffer | ||
| + | * 1μL EcoRI fast digest enzyme | ||
| + | * 1μL SpeI fast digest enzyme | ||
| + | |||
| + | - | ||
| + | |||
| + | * 6μL MQ | ||
| + | * 10μL plasmid in MQ ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100]) | ||
| + | * 2μL Fast digest buffer | ||
| + | * 1μL EcoRI fast digest enzyme | ||
| + | * 1μL SpeI fast digest enzyme | ||
| + | |||
| + | The cups were incubated in a heat block at 37°C for 15 min. followed by adding 4μL 6x loading buffer for gel electroforese. | ||
| + | |||
| + | |||
'''Gel Purification of GVP''' | '''Gel Purification of GVP''' | ||
| Line 10: | Line 39: | ||
* ..mg of gel containing the desired fragment was dissolved in ..μL binding buffer by heating to ..°C for ..min. | * ..mg of gel containing the desired fragment was dissolved in ..μL binding buffer by heating to ..°C for ..min. | ||
* the column was prepared with ..μL | * the column was prepared with ..μL | ||
| + | |||
| + | ===Transporters=== | ||
| + | |||
| + | |||
| + | ==Dry== | ||
===Transporters=== | ===Transporters=== | ||
Revision as of 10:51, 13 July 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
GVP Cluster
Restriction for Assembly
The vector containing GVP cluster was cut with EcoRI and XbaI to create correct ends.
- 6μL MQ
- 10μL plasmid in MQ
- 2μL Fast digest buffer
- 1μL EcoRI fast digest enzyme
- 1μL XbaI fast digest enzyme
The vectors containing promotors [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106] were cut with EcoRI and SpeI.
- 0μL MQ
- 16μL plasmid in MQ ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106])
- 2μL Fast digest buffer
- 1μL EcoRI fast digest enzyme
- 1μL SpeI fast digest enzyme
-
- 6μL MQ
- 10μL plasmid in MQ ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100])
- 2μL Fast digest buffer
- 1μL EcoRI fast digest enzyme
- 1μL SpeI fast digest enzyme
The cups were incubated in a heat block at 37°C for 15 min. followed by adding 4μL 6x loading buffer for gel electroforese.
Gel Purification of GVP
A standard kit for PCR-product purification was used for gel purification
- ..mg of gel containing the desired fragment was dissolved in ..μL binding buffer by heating to ..°C for ..min.
- the column was prepared with ..μL
Transporters
Dry
Transporters
Dry
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