从胰液中提取DNA用于PCR扩增试验的简易方法。

P Müller, R Jesnowski, S Liebe, A Rolfs, M Löhr
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引用次数: 15

摘要

结论:从胰液中制备DNA用于后续的聚合酶链反应(PCR)是困难的,但易于处理。该协议提供了一种简单、快速的解决方案。这种方法可能适用于其他复杂的样品,如唾液、分泌物或粪便洗涤。背景:在所有用于PCR扩增的生物样品中,胰液是最有问题的,因为存在潜在的抑制物质和核酸酶的数量。这就需要一种适合常规诊断PCR的DNA制备程序,因此是高效和安全的。在常规内镜检查中获得的胰液尤其如此。方法:我们在这里描述了一种简单的方法,利用改性苯酚/氯仿直接从天然胰液中提取和沉淀,适用于诊断PCR应用,如癌基因。结果:常规内镜标本可定量制备DNA。DNA也可以从在室温下保存几天的样品中制备出来。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Simple method for DNA extraction from pancreatic juice for PCR amplification assays.

Conclusion: Preparation of DNA from pancreatic juice for subsequent polymerase chain reaction (PCR) is difficult, but manageable. The protocol presented offers a simple and fast solution. This method might be applicable to other complicated samples, such as saliva, would secretions, or stool washings.

Background: Of all the biological samples used for PCR amplification, pancreatic juice is the most problematic because of the presence of potential inhibitory substances and the amount of nucleases. This demands a DNA preparation procedure that is suitable for routine diagnostic PCR, and is therefore efficient and safe. This is particularly true for pancreatic juice obtained during routine endoscopy.

Methods: We describe here a simple method utilizing modified phenol/chloroform extraction and precipitation directly from native pancreatic juice suitable for diagnostic PCR applications, such as oncogenes.

Results: DNA could be prepared in quantitative amounts from routine endoscopic specimens. DNA could also be prepared from samples kept several days at room temperature.

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