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Decontamination of MDA reagents for single cell whole genome amplification.

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.

SUBMITTER: Woyke T

PROVIDER: S-EPMC3197606 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Decontamination of MDA reagents for single cell whole genome amplification.

Woyke Tanja T Sczyrba Alexander A Lee Janey J Rinke Christian C Tighe Damon D Clingenpeel Scott S Malmstrom Rex R Stepanauskas Ramunas R Cheng Jan-Fang JF

PloS one 20111020 10

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, s ...[more]

PMID: 22028825

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