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PLoS One. 2014 May 2;9(5):e96492. doi: 10.1371/journal.pone.0096492. eCollection 2014.

Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment.

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1
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.

Abstract

We have developed a PCR method, coined Déjà vu PCR, that utilizes six nucleotides in PCR with two methyl specific restriction enzymes that respectively digest these additional nucleotides. Use of this enzyme-and-nucleotide combination enables what we term a "DNA diode", where DNA can advance in a laboratory in only one direction and cannot feedback into upstream assays. Here we describe aspects of this method that enable consecutive amplification with the introduction of a 5th and 6th base while simultaneously providing methylation dependent mitochondrial DNA enrichment. These additional nucleotides enable a novel DNA decontamination technique that generates ephemeral and easy to decontaminate DNA.

PMID:
24788618
PMCID:
PMC4008621
DOI:
10.1371/journal.pone.0096492
[Indexed for MEDLINE]
Free PMC Article
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