Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Carretera de Infiesto, 33300 Villaviciosa, Spain.
Anal Biochem. 2010 May 15;400(2):307-9. doi: 10.1016/j.ab.2010.02.010. Epub 2010 Feb 13.
This work describes a simple method for extracting high-quality microbial RNA from fermented milk. Pretreatment consists merely of adding a concentrated solution of sodium citrate to the milk product. The proposed method provides consistently better yields of high-quality RNA than the conventional method independent of the texture of the fermented milk. Furthermore, it can be used with small amounts of starting material, requiring only the use of Eppendorf-size centrifuges-a great advantage for analytical laboratories. The RNA obtained is suitable for the detection of live microorganisms and for transcriptional studies based on quantitative reverse transcription polymerase chain reaction (RT-qPCR).
本工作描述了一种从发酵乳中提取高质量微生物 RNA 的简单方法。预处理只需在乳产品中加入浓的柠檬酸钠溶液。与发酵乳的质地无关,与传统方法相比,该方法提供了始终更好的高质量 RNA 产量。此外,它可以用于少量起始材料,仅需要使用 Eppendorf 大小的离心机-这对分析实验室是一个很大的优势。获得的 RNA 适用于活微生物的检测和基于定量逆转录聚合酶链反应 (RT-qPCR) 的转录研究。