Polysome profiling of oocytes, early development, and cell cycle in mammals

Abstract

Polysome profiling has provided a gold-standard method to precisely assay functional genome readout at a given time point. The wide-spread utilisation of polysome profiling has nevertheless been historically hindered by its intrinsic complexity, time-consuming nature, limited capacity for high throughput adaptation and its requirement for relatively large initial sample sizes. Here, we present Scarce Sample Polysome Profiling (SSP-profiling); method that demonstrates a combination of the sucrose gradient ultracentrifugation in small SW55Ti tubes with the qRT-PCR-based quantification of 18S and 28S rRNAs in fractionated polysome profile. SSP-profiling is suitable for both scarce and conventional sample sizes and is compatible with downstream RNA-seq to identify polysome associated transcripts