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.
Keywords: Polysome; Ribosome fractionation; Sucrose density gradient; Translation; Ultracentrifugation; mRNA.