Qiaamp dneasy mini kit

Total RNA from tissue (>50 mg) was extracted using TRI reagent (Sigma Aldrich, St. Louis, MO) using standard protocol. The quantity and quality of extracted RNA was examined spectrophotometrically. DNA isolation was done from 100mg of tissue samples stored in RNA later solution at -800C degree using QIAamp DNeasy Mini Kit (QIAGEN)and eluted DNA was dissolved in 200µl of AE buffer and stored at 40C for further use. Genomic DNA was also isolated from 100µl of blood following instructions of QIAamp DNeasy Mini Kit (QIAGEN) and eluted DNA was dissolved in 200µl of AE buffer and stored at 40C until further use.

The genetic material was extracted using the QIAampDNeasy Mini Kit (QIAGEN, Germany) that works based on silica gel membrane technology which allowed an efficient recovery ofcomplete DNA from plant tissues. DNA was extracted from each of the samples between 3-5 times.
All PCR amplification runs were performed using an ABI Simpliamp System (Life Technologies, USA). Each amplification reaction contained 1X reaction buffer, 0.1-0.4 μM of each primer (Table S2); 1 U Taq DNA Polymerase (Sinaclon, Iran), 1.5-3 mM MgCl2, 0.20-0.25mM each dATP, dCTP, dGTP, and dTTP (Sinaclon, Iran). Either 2 or 5μ LDNA was added to 15 or 18 μl prepared Master mixes. All PCR reactions were performed based on Tables S3 andS4.