Isolation of high quality RNA from pistachio (Pistacia vera L.) and other woody plants high in secondary metabolites.

ABSTRACT: The quality and quantity of RNA are critical for successful downstream transcriptome-based studies such as microarrays and RNA sequencing (RNA-Seq). RNA isolation from woody plants, such as Pistacia vera, with very high amounts of polyphenols and polysaccharides is an enormous challenge. Here, we describe a highly efficient protocol that overcomes the limitations posed by poor quality and low yield of isolated RNA from pistachio and various recalcitrant woody plants. The key factors that resulted in a yield of 150 μg of high quality RNA per 200 mg of plant tissue include the elimination of phenol from the extraction buffer, raising the concentration of β-mercaptoethanol, long time incubation at 65 °C, and nucleic acid precipitation with optimized volume of NaCl and isopropyl alcohol. Also,

SUBMITTER: Moazzam Jazi M 

PROVIDER: S-EPMC4646865 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature