Decontamination of MDA reagents for single cell whole genome amplification.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 22028825.
- Also identified by DOI 10.1371/journal.pone.0026161 and PMC identifier 3197606.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Single cell genomics is a powerful and increasingly popular tool for studying the genetic make-up of uncultured microbes. A key challenge for successful single cell sequencing and analysis is the removal of exogenous DNA from whole genome amplification reagents. We found that UV irradiation of the multiple displacement amplification (MDA) reagents, including the Phi29 polymerase and random hexamer primers, effectively eliminates the amplification of contaminating DNA. The methodology is quick, simple, and highly effective, thus significantly improving whole genome amplification from single cells.
Medical subject headings
- Artifacts
- DNA Contamination
- Genomics
- Nucleic Acid Amplification Techniques
- Single-Cell Analysis
- Ultraviolet Rays