A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples
Wang,Hang; Qi,Jinfeng; Xiao,Derong; Wang,Zhibao; Tian,Kun
2017
发表期刊SOIL BIOLOGY & BIOCHEMISTRY
ISSN0038-0717
卷号106页码:109-118
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.
关键词Real-time Pcr Environmental-samples Extraction Sediment Quantification Optimization Assay Pcr Inhibition Dilution Dna Extraction Microbe Humic Acid
收录类别SCI
语种英语
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文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/54961
专题植物化学与西部植物资源持续利用国家重点实验室
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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.
APA 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.
MLA Wang,Hang,et al."A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples".SOIL BIOLOGY & BIOCHEMISTRY 106(2017):109-118.
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