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Pre-lysis washing improves DNA extraction from a forest soil
He, JZ; Xu, ZH; Hughes, J
2005
Source PublicationSOIL BIOLOGY & BIOCHEMISTRY
Volume37Issue:12Pages:2337-2341
AbstractA pre-lysis buffer washing procedure was introduced to DNA extraction from a forest soil with high organic matter and iron oxide contents. Sodium phosphate of 0.1 M (pH 7.5) was used as a buffer to wash soil samples when subsequent lysis buffer was phosphate, and 20 mM EDTA (pH 7.5) was used when subsequent lysis buffer included EDTA. Initial experiments were not successful because the DNA extracts could not be amplified by polymerase chain reaction (PCR). The consideration of introducing a pre-lysis washing procedure was based on the idea that the washing should promote soil dispersion and homogeneity, decrease DNA adsorption by soil components (e.g. iron oxides), and remove covalent cations and those easily-dissolving organic compounds from the soil samples. Results revealed that humic substance content decreased by 31%, but DNA yield increased by 24% in the DNA extracts of the pre-lysis washing procedures, compared to the non-washing procedures. DNA extracted by the pre-washing procedure needed less purification for subsequent 18S and 16S rDNA PCR amplifications. It was recommended that the pre-lysis buffer washing should be used for DNA extraction from those difficult environmental samples, such as the forest soil with high contents of organic matter and iron oxides. (c) 2005 Elsevier Ltd. All rights reserved.
Department土壤环境科学实验室
KeywordDna Extraction Buffer Washing Forest Soil Humic Substance Iron Oxides Polymerase Chain Reaction
DOI10.1016/j.soilbio.2005.04.016
WOS Research AreaAgriculture
WOS IDWOS:000234327700021
Citation statistics
Cited Times:49[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.rcees.ac.cn/handle/311016/23352
Collection土壤环境科学实验室
Recommended Citation
GB/T 7714
He, JZ,Xu, ZH,Hughes, J. Pre-lysis washing improves DNA extraction from a forest soil[J]. SOIL BIOLOGY & BIOCHEMISTRY,2005,37(12):2337-2341.
APA He, JZ,Xu, ZH,&Hughes, J.(2005).Pre-lysis washing improves DNA extraction from a forest soil.SOIL BIOLOGY & BIOCHEMISTRY,37(12),2337-2341.
MLA He, JZ,et al."Pre-lysis washing improves DNA extraction from a forest soil".SOIL BIOLOGY & BIOCHEMISTRY 37.12(2005):2337-2341.
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