PLoS ONE · 2011 · 212 citations · 12 references
Life SciencesMicrobiologyMolecular BiologyGenetic EngineeringGenomicsMicrobial GeneticsDna SequencingGenome SequencingPolymerase Chain ReactionSingle-cell GenomicsDisplacement AmplificationMultiple Displacement AmplificationSingle-cell Dna SequencingSingle-cell Genome SequencingSingle-cell Whole-genome SequencingStrand Displacement AmplificationUv InactivationWhole-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.
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UV-induced mutation hotspots occur at DNA damage hotspots
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