|A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples|
|Wang,Hang; Qi,Jinfeng; Xiao,Derong; Wang,Zhibao; Tian,Kun
|发表期刊||SOIL BIOLOGY & BIOCHEMISTRY
|摘要||Quantitative real-time PCR (qPCR) analysis for accurate quantification of targeted microbial genes is compromised by the presence of co-extracted inhibitors from soil samples. Dilution of DNA extracts is a commonly-used method to reduce levels of inhibition. However, the applications of dilution method are mostly empirical, and need to be further elaborated. Here, we propose a dilution model to re-evaluate dilution as a method to eliminate qPCR inhibition. We found that DNA extracts without dilution or with a minor dilution (e.g., 10-fold) resulted in ciPCR inhibition for most of studied soils. However, excessive dilution (e.g., 200- or 400-fold) caused an overestimation of the quantified gene cdpy numbers. Only under a moderate dilution range could qPCR inhibition be efficiehtly eliminated, which has been well captured by our proposed dilution model. The pre-testing of qPCR inhibition for determining the appropriate dilution range for extracted DNA samples aids accurate quantification of nucleic acids in soils. (C) 2016 Elsevier Ltd. All rights reserved.|
Wang,Hang,Qi,Jinfeng,Xiao,Derong,et al. A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples[J]. SOIL BIOLOGY & BIOCHEMISTRY,2017,106:109-118.
Wang,Hang,Qi,Jinfeng,Xiao,Derong,Wang,Zhibao,&Tian,Kun.(2017).A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples.SOIL BIOLOGY & BIOCHEMISTRY,106,109-118.
Wang,Hang,et al."A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples".SOIL BIOLOGY & BIOCHEMISTRY 106(2017):109-118.