CHELEX-100 AS A MEDIUM FOR SIMPLE EXTRACTION OF DNA FOR PCR-BASED TYPING FROM FORENSIC MATERIAL
CHELEX-100 AS A MEDIUM FOR SIMPLE EXTRACTION OF DNA FOR PCR-BASED TYPING FROM FORENSIC MATERIAL
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DOI：
10.2144/000114018
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发表时间：
1991-04-01
期刊：
影响因子：
2.7
通讯作者：
HIGUCHI, R
中科院分区：
文献类型：
--
作者：
WALSH, PS;METZGER, DA;HIGUCHI, R
已经开发了利用Chelex（R）100螯合树脂的程序，用于从法医型样品中提取DNA以用于PCR。 该方法简单、快速，不涉及有机溶剂，对大多数类型的样品不需要多管转移。 使用Chelex 100从精液和非常小的血迹中提取DNA与使用蛋白酶K和酚-氯仿提取一样有效或更有效。 从血迹中提取的DNA似乎不太容易含有PCR抑制剂时，通过这种方法制备。 Chelex方法已用于HLA DQ-α基因座的扩增和分型，以获得许多不同类型样本的DQ-α基因型，包括全血、血迹、精斑、口腔拭子、毛发和性交后样本。 一致性研究的结果，其中DQ-α基因型的84个样品制备使用Chelex或使用传统的酚-氯仿提取进行了比较。 使用两种不同的提取方法获得的基因型是相同的所有样品测试。
Procedures utilizing Chelex(R) 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube trasfers for most types of samples. The extraction of DNA from semen and very small bloodstains using Chelex 100 is as efficient or more efficient than using proteinase K and phenol-chloroform extraction. DNA extracted from bloodstains seems less prone to contain PCR inhibitors when prepared by this method. The Chelex method has been used with amplification and typing at the HLA DQ-alpha locus to obtain the DQ-alpha genotypes of many different types of samples, including whole blood, bloodstains, seminal stains, buccal swabs, hair and post-coital samples. The results of a concordance study are presented in which the DQ-alpha genotypes of 84 samples prepared using Chelex or using conventional phenol-chloroform extraction are compared. The genotypes obtained using the two different extraction methods were identical for all samples tested.