Direct polymerase chain reaction from whole blood without DNA isolation
Direct polymerase chain reaction from whole blood without DNA isolation
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DOI：
10.1258/0004563001899726
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发表时间：
2000-09-01
影响因子：
2.2
通讯作者：
Kato, S
中科院分区：
文献类型：
--
作者：
Nishimura, N;Nakayama, T;Kato, S
血液和其他动物体液含有多种抑制聚合酶链式反应 (PCR) 的物质，因此在 PCR 之前通常需要分离 DNA。我们开发了一种新型试剂混合物，可以有效抑制这些抑制物质，使得 PCR 不需要从血液中分离 DNA。当该试剂包含在 PCR 混合物中时，可以直接从用各种抗凝剂或 PCR 抑制物质处理的人体血液样本中有效地扩增 β-珠蛋白基因的 DNA 片段。我们通过使用各种 PCR 引物组检查大量血液样本，证实了这种鸡尾酒的有用性。除了新鲜血液之外，该方法还可以对在 4°C - 20°C 或 - 80°C 下保存至少 1 年的血液样本进行 PCR 扩增。
Blood and other animal fluids contain a variety of substances that inhibit the polymerase chain reaction (PCR), so that isolation of DNA is generally necessary prior to PCR. We have developed a novel reagent cocktail that effectively suppresses these inhibitory substances and makes DNA isolation from blood unnecessary for PCR. When this reagent was included in the PCR mixture, DNA Fragments of the beta-globin gene could be efficiently amplified directly from human blood samples treated with various anticoagulants or PCR-inhibitory substances. We confirmed the usefulness of this cocktail by examining a large number of blood samples with various PCR primer sets. Tn addition to fresh blood, this method enabled PCR amplification from blood samples stored at 4 degrees C. - 20 degrees C or - 80 degrees C for a minimum of 1 year.