Decontamination of MDA reagents for single cell whole genome amplification

Autor: Alexander Sczyrba, Tanja Woyke, Damon Tighe, Jan Fang Cheng, Scott Clingenpeel, Ramunas Stepanauskas, Rex R. Malmstrom, Janey Lee, Christian Rinke
Rok vydání: 2011
Předmět:
Time Factors
Ultraviolet Rays
Applied Microbiology
lcsh:Medicine
Genomics
Computational biology
Bacillus Phages
DNA-Directed DNA Polymerase
Biology
Microbiology
Microbial Ecology
03 medical and health sciences
Environmental Biotechnology
Genomic library
Genome Sequencing
lcsh:Science
Genome Evolution
030304 developmental biology
DNA Primers
Whole Genome Amplification
0303 health sciences
Multidisciplinary
Escherichia coli K12
Ecology
030306 microbiology
lcsh:R
Microbial Mutation
Multiple displacement amplification
MALBAC
Microbial Growth and Development
Dose-Response Relationship
Radiation

Nucleic acid amplification technique
Sequence Analysis
DNA

Biodiversity
DNA Contamination
Molecular biology
Single cell sequencing
Microbial Evolution
lcsh:Q
Indicators and Reagents
Metagenomics
Single-Cell Analysis
Artifacts
Applications of PCR
Nucleic Acid Amplification Techniques
Research Article
Biotechnology
Zdroj: PLoS ONE
PLoS ONE, Vol 6, Iss 10, p e26161 (2011)
ISSN: 1932-6203
Popis: 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.
Databáze: OpenAIRE