A re-evaluation of dilution for eliminating PCR inhibition in soil DNA samples

doi:10.1016/j.soilbio.2016.12.011

	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
ISSN	0038-0717
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
学科领域	Soil Science
DOI	10.1016/j.soilbio.2016.12.011
收录类别	SCI
语种	英语
引用统计
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
推荐引用方式 GB/T 7714	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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