Optimization of RNA isolation from Brittle Leaf Disease affected date palm leaves and construction of a subtractive cDNA library.
A simple and efficient method was described here for the isolation of high-quality RNA from date palm leaves affected with Brittle Leaf Disease (BLD) and containing high amount of phenolic compounds. The procedure was based on the use of a non-ionic detergent Nonidet-P40 (NP-40), Polyvinylpyrrolidone (PVP), and beta-mercaptoethanol in the extraction buffer in order to isolate cytoplasmic RNA and to prevent the oxidation of phenolic compounds. This method allowed the isolation of intact RNA, suitable for cDNA synthesis and library construction. Differential screening of the subtractive cDNA library from affected leaf RNA led to the identification of some BLD-induced genes.
Saïdi
Mohammed Najib
MN
Laboratoire des Biotechnologies Végétales Appliquées à l'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, Route Soukra Km 4, Sfax, Tunisia. saidimn@yahoo.fr
Gargouri-Bouzid
Radhia
R
Rayanni
Mariem
M
Drira
Noureddine
N
D028023:Arecaceae / Q000235:genetics*; D005419:Flavonoids / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D008623:Mercaptoethanol / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D017830:Octoxynol; D010636:Phenols / Q000378:metabolism; D010935:Plant Diseases / Q000235:genetics*; D018515:Plant Leaves / Q000235:genetics*; D011092:Polyethylene Glycols / Q000737:chemistry; D059808:Polyphenols; D011205:Povidone / Q000737:chemistry; D018749:RNA, Plant / Q000302:isolation & purification* / Q000378:metabolism
D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols; D018749:RNA, Plant; D011092:Polyethylene Glycols; D008623:Mercaptoethanol; D017830:Octoxynol; C010615:Nonidet P-40; D011205:Povidone