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 advan...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096492&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850191708830040064 |
|---|---|
| author | Kevin J McKernan Jessica Spangler Lei Zhang Vasisht Tadigotla Stephen McLaughlin Jason Warner Amir Zare Richard G Boles |
| author_facet | Kevin J McKernan Jessica Spangler Lei Zhang Vasisht Tadigotla Stephen McLaughlin Jason Warner Amir Zare Richard G Boles |
| author_sort | Kevin J McKernan |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-8f03bc36dff14e60b4fbb2011b8f378d |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-8f03bc36dff14e60b4fbb2011b8f378d2025-08-20T02:14:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0195e9649210.1371/journal.pone.0096492Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment.Kevin J McKernanJessica SpanglerLei ZhangVasisht TadigotlaStephen McLaughlinJason WarnerAmir ZareRichard G BolesWe 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096492&type=printable |
| spellingShingle | Kevin J McKernan Jessica Spangler Lei Zhang Vasisht Tadigotla Stephen McLaughlin Jason Warner Amir Zare Richard G Boles Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment. PLoS ONE |
| title | Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment. |
| title_full | Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment. |
| title_fullStr | Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment. |
| title_full_unstemmed | Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment. |
| title_short | Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment. |
| title_sort | expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096492&type=printable |
| work_keys_str_mv | AT kevinjmckernan expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT jessicaspangler expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT leizhang expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT vasishttadigotla expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT stephenmclaughlin expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT jasonwarner expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT amirzare expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment AT richardgboles expandedgeneticcodesinnextgenerationsequencingenabledecontaminationandmitochondrialenrichment |