Amplified insert assembly: An optimized approach to standard assembly of BioBrick TM genetic circuits

Michael A. Speer, Tom L. Richard

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

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%.

Original languageEnglish (US)
Article number17
JournalJournal of Biological Engineering
Volume5
DOIs
StatePublished - Dec 16 2011

All Science Journal Classification (ASJC) codes

  • Environmental Engineering
  • Biomedical Engineering
  • Molecular Biology
  • Cell Biology

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