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J Biol Eng. 2011 Dec 16;5(1):17.

Amplified insert assembly: an optimized approach to standard assembly of BioBrickTM genetic circuits.

Speer MA, Richard TL.

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Department of Agricultural and Biological Engineering, The Pennsylvania State University, 249 Agricultural Engineering Bldg,, University Park, PA 16802, USA. trichard@psu.edu.

Abstract

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%.

PMID:: 22176971; [PubMed - in process]
PMCID:: PMC3287150

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