GSM3473832: BB7; Solanum lycopersicum; RNA-Seq
Two grams of tissue per sample were ground in liquid nitrogen and 10mL of the RNA extraction buffer (CTAB 2% v/v, PVP 2%v/v, 100mMTris pH 8, 2MNaCl, 25mM EDTA, 0.5 g/L spermidine, 10mM β-mercaptoethanol) were added. The samples were immediately incubated for 5min at 65◦C. Two extractions with one equal volume of chloro- form:isoamyl alchohol (24:1, v/v) followed by centrifugation at 4000 rpm for 45 min at 4◦C were performed. The supernatant was recovered and 1/10 volume of 1MKOAc was added followed by centrifugation at 4000 rpm for 20 min at 4◦C. The super- natant was collected and 1/4 volume of 10M LiCl was added. Samples were incubated overnight at−20◦C and then centrifuged at 4000 rpm for 45 min at 4◦C. The supernatant was discarded and the RNA pellet was further purified using the RNeasy Plant Mini Kit (Qiagen®). DNAse treatment (RNase-Free DNase Set,Qiagen®) was done in column during the purification step. The RNA was resuspended in 35µL of nuclease-free water. The RNA concentration and purity were measured using NanoDrop 2000c Spectrophotometer (Thermo Scientific, Inc.). The RNA integrity was checked by agarose gel electrophoresis. Libraries were prepared using the Illumina TruSeq RNA Sample Preparation Kit v2 (Illumina, CA).
GEO Accession
GSM3473832