Decontamination of MDA reagents for single cell whole genome amplification.
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.
Department of Energy Joint Genome Institute, Walnut Creek, California, United States of America.
Tanja
T
Woyke
Alexander
A
Sczyrba
Janey
J
Lee
Christian
C
Rinke
Damon
D
Tighe
Scott
S
Clingenpeel
Rex
R
Malmstrom
Ramunas
R
Stepanauskas
Jan-Fang
JF
Cheng
D016477:Artifacts; D017103:Bacillus Phages / Q000201:enzymology; D059018:DNA Contamination; D017931:DNA Primers / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004307:Dose-Response Relationship, Radiation; D048168:Escherichia coli K12 / Q000166:cytology / Q000235:genetics; D023281:Genomics / Q000379:methods*; D007202:Indicators and Reagents / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D017422:Sequence Analysis, DNA; D059010:Single-Cell Analysis / Q000379:methods*; D013997:Time Factors; D014466:Ultraviolet Rays
D017931:DNA Primers; D007202:Indicators and Reagents; D004259:DNA-Directed DNA Polymerase