A Rapid and Efficient Method for Purifying High Quality Total RNA from Peaches (Prunus persica) for Functional Genomics Analyses

Abstract

Prunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica). Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilum is ideal for isolating high quality RNA from the fruits of Prunus persica. This RNA may be used for many functional genomic based experiments such as RT-PCR and the construction of large-insert cDNA libraries.

Reconstitution of a 10-gene pathway for synthesis of the plant alkaloid dihydrosanguinarine in Saccharomyces cerevisiae

“…RNA extraction from root or stems of the P. somniferum (opium poppy) cultivar Bea's Choice was performed using a modified CTAB method 51 and the cDNA libraries were constructed using the TruSeq Stranded mRNA Sample Prep Kit (Illumina). Illumina GA sequencing was performed with 7 pmol per library with one lane in the flow cell.…”

Development of ChillPeach genomic tools and identification of cold-responsive genes in peach fruit

“…Total RNA was isolated from 4 g of pooled mesocarp tissue of three fruit in each treatment using the method described by Meisel et al (2005). For both libraries, 10 lg RNA of M, TR and R, and 30 lg RNA of CS1, CSR-1, CS-2 and CSR-2 were pooled and used for cDNA synthesis.…”

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

“…The total RNA was isolated in triplicate for each sample and yields ranged from 0.67 to 1.09 μg per mg of fresh weight, which was considerably high in terms of the quantity and quality of the RNA relative to other reports as presented in Fig. 1b and Table 2 (Gasic et al 2004;Meisel et al 2005;Provost et al 2007;Ghawana et al 2011).…”