Title |
Expanded Genetic Codes in Next Generation Sequencing Enable Decontamination and Mitochondrial Enrichment
|
---|---|
Published in |
PLOS ONE, May 2014
|
DOI | 10.1371/journal.pone.0096492 |
Pubmed ID | |
Authors |
Kevin J. McKernan, Jessica Spangler, Lei Zhang, Vasisht Tadigotla, Stephen McLaughlin, Jason Warner, Amir Zare, Richard G. Boles |
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. |
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