fulltext.study @t Gmail

Simplified gene synthesis: A one-step approach to PCR-based gene construction

Paper ID Volume ID Publish Year Pages File Format Full-Text
25711 43593 2006 8 PDF Available
Title
Simplified gene synthesis: A one-step approach to PCR-based gene construction
Abstract

PCR-based gene synthesis conventionally requires two steps: first, all overlapping oligonucleotides are assembled by self-priming; then an additional pair of primers is used to amplify the full-length gene product. Here we propose a simplified method of gene synthesis which combines these two steps into one. We have found that the efficiency of this one-step method, which we term “Simplified Gene Synthesis”, is affected by multiple parameters of the PCR reactions. In particular, the choice of polymerase is critical for successful one-step assembly. Other important factors include the concentration of assembly oligonucleotides and amplification primers. Moreover, we offer a general method to estimate, given a known mutation rate, how many clones should be sequenced in order to be confident of obtaining at least one correct gene product. Having determined the accuracy of gene products synthesized under optimal conditions with Simplified Gene Synthesis, we show that our estimation works well. Overall, the simplified gene synthesis provides an easier and more efficient approach to gene synthesis, providing a further step towards the future goal of generalized automation for this process.

Keywords
SGS, simplified gene synthesis; AO, assembly oligonucleotides; AP, amplification primer; GC, guanine and cytosineGene synthesis; PCR-based; KOD polymerase; Optimal concentration; Accuracy
First Page Preview
Simplified gene synthesis: A one-step approach to PCR-based gene construction
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 124, Issue 3, 25 July 2006, Pages 496–503
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us