Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment.
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.
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Kevin J
KJ
McKernan
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Jessica
J
Spangler
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Lei
L
Zhang
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Vasisht
V
Tadigotla
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Stephen
S
McLaughlin
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Jason
J
Warner
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Amir
A
Zare
Courtagen Life Sciences, Woburn, Massachusetts, United States of America.
Richard G
RG
Boles
D003062:Codon / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D019175:DNA Methylation; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000378:metabolism; D005815:Genetic Code; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008957:Models, Genetic; D009711:Nucleotides / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
D003062:Codon; D004272:DNA, Mitochondrial; D009711:Nucleotides; D004247:DNA; D004262:DNA Restriction Enzymes