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评价五种基于柱的分离试剂盒及其从人乳中提取 miRNA 的能力。

Evaluation of five column-based isolation kits and their ability to extract miRNA from human milk.

机构信息

Division of Inflammation and Infection, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden.

Örebro University Food and Health Programme, School of Medical Sciences, Örebro University, Örebro, Sweden.

出版信息

J Cell Mol Med. 2021 Aug;25(16):7973-7979. doi: 10.1111/jcmm.16726. Epub 2021 Jun 27.

Abstract

MicroRNA can be found in various body fluids, including breast milk. MicroRNA may be transferred from mother to infant via breast milk and potentially regulate the development of the infant's immune system on a post-transcriptional level. This study aimed to determine the microRNA extraction efficiency of five RNA extraction kits from human skim milk samples. Their efficiency was determined by comparing microRNA concentrations, total RNA yield and purity. Furthermore, hsa-miR-148a-3p expression and the recovery of an exogenous control, cel-miR-39-3p, were quantified using qPCR. Each kit extracted different amounts of microRNA and total RNA, with one kit tending to isolate the highest amount of both RNA species. Based on these results, the extraction kit ReliaPrep™ miRNA Cell and Tissue Miniprep System from Promega was found to be the most appropriate kit for microRNA extraction from human skim milk. Moreover, further research is needed to establish a standardized protocol for microRNA extraction from breast milk.

摘要

微小 RNA 存在于各种体液中,包括母乳。微小 RNA 可能通过母乳从母亲转移到婴儿,并在转录后水平上潜在地调节婴儿免疫系统的发育。本研究旨在确定五种 RNA 提取试剂盒从人脱脂乳样品中提取微小 RNA 的效率。通过比较微小 RNA 浓度、总 RNA 产量和纯度来确定它们的效率。此外,使用 qPCR 定量检测 hsa-miR-148a-3p 的表达和外源性对照 cel-miR-39-3p 的回收率。每个试剂盒提取的微小 RNA 和总 RNA 量不同,其中一个试剂盒倾向于分离两种 RNA 物质的最高量。基于这些结果,发现 Promega 的 ReliaPrep™ miRNA Cell and Tissue Miniprep System 试剂盒最适合从人脱脂乳中提取微小 RNA。此外,需要进一步的研究来建立从母乳中提取微小 RNA 的标准化方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8739/8358847/9d67b8701513/JCMM-25-7973-g002.jpg

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