Impact of Contaminating DNA in Whole-Genome Amplification Kits Used for Metagenomic Shotgun Sequencing for Infection Diagnosis.

摘要: ABSTRACT Whole-genome amplification (WGA) is a useful tool for of very small quantities DNA many uses, including metagenomic shotgun sequencing infection diagnosis. Depending on the application, background from WGA kits can be problematic. Three were tested their utility in metagenomics approach to identify pathogens sonicate fluid comprised biofilms and other materials dislodged surfaces explanted prosthetic joints using sonication. The Illustra V2 Genomiphi, single cell Qiagen REPLI-g used test identical samples. Variations number reads, genera identified background, reads known present samples observed between kits. These results then compared those obtained with library preparation without prior an NEBNext Ultra II paired-end kit, which requires amount input DNA. This also resulted presence contaminant bacterial yielded fewer pathogens. findings highlight impact that kit selection have analysis low-biomass importance careful consideration implications these tools.