An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development.

ABSTRACT:

Background

Accuracy in quantitative real-time RT-PCR is dependent on high quality RNA, consistent cDNA synthesis, and validated stable reference genes for data normalization. Reference genes used for normalization impact the results generated from expression studies and, hence, should be evaluated prior to use across samples and treatments. Few statistically validated reference genes have been reported in grapevine. Moreover, success in isolating high quality RNA from grapevine tissues is typically limiting due to low pH, and high polyphenolic and polysaccharide contents.

Results

We describe optimization of an RNA isolation procedure that compensates for the low pH found in grape berries and improves the ability of the RNA to precipitate. This procedure was tested on pericar

SUBMITTER: Reid KE 

PROVIDER: S-EPMC1654153 | biostudies-literature | 2006 Nov

REPOSITORIES: biostudies-literature