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.
MeSH Terms
Artifacts
Bacillus Phages/enzymology
DNA Contamination
DNA Primers/genetics,metabolism
DNA-Directed DNA Polymerase/metabolism
Dose-Response Relationship, Radiation
Escherichia coli K12/cytology,genetics
Genomics/methods
Indicators and Reagents/metabolism
Nucleic Acid Amplification Techniques/methods
Sequence Analysis, DNA
Single-Cell Analysis/methods
Time Factors
Ultraviolet Rays
Chemicals
DNA Primers
Indicators and Reagents
DNA-Directed DNA Polymerase
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Woyke Tanja
Department of Energy Joint Genome Institute, Walnut Creek, California, United States of America.
Sczyrba Alexander
Lee Janey
Rinke Christian
Tighe Damon
Clingenpeel Scott
Malmstrom Rex
Stepanauskas Ramunas
Cheng Jan-Fang
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