Amplified insert assembly: an optimized approach to standard assembly of BioBrickTM genetic circuits.
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ABSTRACT:
SUBMITTER: Speer MA
PROVIDER: S-EPMC3287150 | biostudies-literature | 2011 Dec
REPOSITORIES: biostudies-literature
Publications
Amplified insert assembly: an optimized approach to standard assembly of BioBrickTM genetic circuits.
Speer Michael A MA Richard Tom L TL
Journal of biological engineering 20111216 1
A modified BioBrick™ assembly method was developed with higher fidelity than current protocols. The method utilizes a PCR reaction with a standard primer set to amplify the inserted part. Background colonies are reduced by a combination of dephosphorylation and digestion with DpnI restriction endonuclease to reduce vector and insert background respectively. The molar ratio of the insert to vector in the ligation was also optimized, with the accuracy of the transformed construct approaching 100%. ...[more]
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