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Team:Hawaii/Initial BioBrick Plasmid Extraction From Filter Paper
From 2008.igem.org
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==Protocol== | ==Protocol== | ||
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'''BioBricks parts used:''' | '''BioBricks parts used:''' | ||
* BBa_E0040 GFP biobrick (spot 7E on plate 1001) | * BBa_E0040 GFP biobrick (spot 7E on plate 1001) | ||
Revision as of 19:24, 10 June 2008
Protocol
BioBricks parts used:
- BBa_E0040 GFP biobrick (spot 7E on plate 1001)
- BBa_J04430 IPTG-induced GFP device (spot 4H on plate 1004)
- BBa_I13522 constitutive GFP device (spot 8A on plate 1002)
Paper punch
- Warmed 5 μl aliquots of TE buffer (pH 8.0) to 50C in 0.5 ml PCR tubes
- Slid cutting mat under page in binder with desired part
- Pressed down w/ punch tool
- Ejected punch spot in TE buffer tube
Transformation
- Soaked spots 20 min in warmed TE buffer
- Thawed competent DH5α cells on ice. Chilled empty 2 ml eppendorf tubes on ice
- Added 2 μl TE + DNA solution to 50 μl thawed cells
- Incubated 30 min on ice
- Heat shocked in 42C water bath for 60 sec
- Incubated 2 min on ice
- Added 200 μl SOC
- Incubated for 2 hr at 37C with shaking (235 rpm)
- Plated 200 μl 1/1, 1/10, 1/100 dilutions on LB+amp
- Incubated overnight at 37C.
Protocol adapted from Registry of Standard Biological Parts, [http://partsregistry.org/wiki/index.php/Help:IGEM_08_DNA_distribution Spring 2008 DNA Distribution].
