Poria Dipak Kumar, Ray Partho Sarothi
Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India.
Bio Protoc. 2017 Mar 20;7(6). doi: 10.21769/BioProtoc.2192.
Polysome analysis is a method to separate mRNAs from a cell into actively translating and non-translating fractions depending on their association with polysomes. By this protocol, cell lysates are fractionated by sucrose density gradient ultracentrifugation. Free mRNA fraction and various ribosomal fractions, such as 40S, 60S, monosomes and polysomes are collected by fractionation. Association of particular mRNAs with these fractions is detected by reverse transcription - PCR to investigate the translational state of the mRNA.
多核糖体分析是一种根据mRNA与多核糖体的结合情况，将细胞中的mRNA分离为活跃翻译部分和非翻译部分的方法。通过该方案，细胞裂解物通过蔗糖密度梯度超速离心进行分级分离。通过分级收集游离mRNA部分以及各种核糖体部分，如40S、60S、单核糖体和多核糖体。通过逆转录-聚合酶链反应检测特定mRNA与这些部分的结合情况，以研究mRNA的翻译状态。
