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		+	== Bio-Brick Verification ==

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Revision as of 22:05, 26 October 2007

Contents 1 Experimentation 1.1 Protocols 1.2 Assembly Line Flow 1.3 Bio-Brick Creation 1.4 Bio-Brick Verification

Experimentation

Protocols

For reliability, a majority of our actual experimentation has followed protocols provided by the supplier of relevant projects, including enzymes ([http://www.neb.com/ NEB]) and kits ([http://www.qiagen.com/ Qiagen]). The [http://www.qiagen.com/ Qiagen] maxiprep kit was replaced with a [http://www.bio-rad.com/ Bio-Rad] kit to obtain higher yields with lower processing times.

Please consult the relevant vendors for protocol documents.

Assembly Line Flow

Bio-Brick Creation

Design Primers

PCR

Run Gel

- Check Primer concentration
- Check Primer design

Gel Extraction and determine concentrations

Digestion with Restriction Enzymes

PCR purification and determine concentrations

Ligation

Transform and pick colonies

Miniprep and determine concentrations

Restriction Map

Retransform with chosen plasmid

Maxiprep and determine concentraions

Sequence

Bio-Brick Verification