Open AccessPolysome Analysis
Author(s) -
Dipak Kumar Poria,
Partho Sarothi Ray
Publication year - 2017
Publication title -
bio-protocol
Language(s) - English
Resource type - Journals
ISSN - 2331-8325
DOI - 10.21769/bioprotoc.2192
Subject(s) - polysome , messenger rna , ribosomal rna , ultracentrifuge , sucrose gradient , ribosome , fractionation , chemistry , microbiology and biotechnology , cell fractionation , biology , biochemistry , rna , chromatography , gene , enzyme
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.