Team:Wash U/Protocol
From 2009.igem.org
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=='''BioBrick Assembly'''== | =='''BioBrick Assembly'''== | ||
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- | :BioBrick Assembly is a standard protocol for combining two BioBrick parts and positioning them in a destination plasmid. Specific cut sites for standard restriction enzymes are utilized to simplify the protocol and include EcoR1-HF, Xba1, Spe1 and Pst1. The assembly consists of two major parts: digestion and ligation. Note that BioBrick assembly requires purified, isolated DNA for all parts involved (upstream, downstream and destination plasmid). To view the full Biobrick Assembly manual, please click [http://ginkgobioworks.com/support/ here]. | + | :BioBrick Assembly is a standard protocol for combining two BioBrick parts and positioning them in a destination plasmid. [http://www.example.com link title]Specific cut sites for standard restriction enzymes are utilized to simplify the protocol and include EcoR1-HF, Xba1, Spe1 and Pst1. The assembly consists of two major parts: digestion and ligation. Note that BioBrick assembly requires purified, isolated DNA for all parts involved (upstream, downstream and destination plasmid). To view the full Biobrick Assembly manual, please click [http://ginkgobioworks.com/support/ here]. |
'''Materials''' | '''Materials''' | ||
* NEBuffer 2 | * NEBuffer 2 | ||
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* purified destination plasmid | * purified destination plasmid | ||
'''Procedures''' | '''Procedures''' | ||
- | # Begin | + | # Begin by thawing all DNA along with NEBuffer 2 and BSA. |
- | + | # Label three separate PCR tubes upstream, downstream, and destination plasmid. To each tube 500ng of respective DNA and dilute to 42.5 uL. Add 5 uL of NEBuffer 2 and 0.5 uL of BSA to each tube. | |
- | # | + | # Add 1 uL of the first appropriate enzyme to each tube (see [http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.jpg chart] for correct enzyme. |
[[Team:Wash_U/Protocol#Procedures|Back To Top]] | [[Team:Wash_U/Protocol#Procedures|Back To Top]] | ||
Revision as of 15:32, 10 July 2009