Method to isolate polyribosomal mRNA from scarce samples such as mammalian oocytes and early embryos.

ABSTRACT:

Background

Although the transcriptome of minute quantities of cells can be profiled using nucleic acid amplification techniques, it remains difficult to distinguish between active and stored messenger RNA. Transcript storage occurs at specific stages of gametogenesis and is particularly important in oogenesis as stored maternal mRNA is used to sustain de novo protein synthesis during the early developmental stages until the embryonic genome gets activated. In many cases, stored mRNA can be several times more abundant than mRNA ready for translation. In order to identify active mRNA in bovine oocytes, we sought to develop a method of isolating very small amounts of polyribosome mRNA.

Results

The proposed method is based on mixing the extracted oocyte cytoplasm with a preparat

SUBMITTER: Scantland S 

PROVIDER: S-EPMC3055227 | biostudies-literature | 2011 Feb

REPOSITORIES: biostudies-literature