Assessment of the quality and quantity of genomic DNA recovered from canine blood samples by three different extraction methods
Assessment of the quality and quantity of genomic DNA recovered from canine blood samples by three different extraction methods
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DOI：
10.1016/j.rvsc.2007.09.009
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发表时间：
2008-08-01
影响因子：
2.4
通讯作者：
Ollier, William E. R.
中科院分区：
文献类型：
--
作者：
Clements, Dylan N.;Wood, Shona;Ollier, William E. R.
基因组DNA （gDNA）提取的理想方法应该是从原始样品源中回收大量纯的、完整的gDNA，同时减少下游过程抑制剂的共提取。采用三种不同的方法（硅胶柱法、苯酚-氯仿法和改进的盐沉淀法）提取经乙二胺四乙酸处理的犬血和凝血标本。苯酚-氯仿和改良盐沉淀法对EDTA保存的血液中gDNA的相对回收率相似，但对凝血中gDNA的回收率较低。分光光度计测量苯酚-氯仿为基础的提取倾向于高估从提取中回收的gDNA的数量，并且与PCR抑制剂的更大的共提取有关。无论样品类型或定量方法如何，二氧化硅柱法都能以相同的效率、纯度和完整性回收gDNA。(C) 2007 Elsevier Ltd.版权所有。
The ideal method for genomic DNA (gDNA) extraction should recover high quantities of pure, integral gDNA from the original sample source with minimal co-extraction of inhibitors of downstream processes. Canine ethylenediamine tetra-acetic acid (EDTA) treated and clotted blood samples were extracted by three different methods (a silica column method, a phenol-chloroform method and a modified salt precipitation method). Phenol-chloroform and modified salt precipitation based extractions demonstrated similar relative recovery of gDNA with EDTA preserved blood, but were less efficient at recovering gDNA from clotted blood. Spectrophotometer measurement of phenol-chloroform based extractions tended to overestimate the quantity of gDNA recovered from extractions, and was associated with the greater co-extraction of PCR inhibitors. The silica column method recovered gDNA with equal efficiency, purity and integrity irrespective of the sample type or method of quantification. (C) 2007 Elsevier Ltd. All rights reserved.