GSM1368809: FLM_untreated_RNA-seq; Tuber melanosporum; RNA-Seq
Total RNA from FLM and FB was extracted as described previously (Montanini et al. 2003) and purified with the RNeasy Plant Mini kit and on-column DNase I digestion (QIAGEN) following the manufacturer’s instructions. RNA integrity was verified by a Bioanalyzer: RIN scores of 7.0 and 6.5 for FB and FLM, respectively. RNA was quantified with the Qubit RNA BR Assay and 1 μg of purified RNA was utilized as starting material for library construction, which was carried out with the Illumina TruSeq RNA Sample Preparation kit following the manufacturer's instructions. Libraries were sequenced using 50-bp single-end reads on the HiSeq 2000 System (Illumina). Tissues were grinded in liquid nitrogen, extracted in the extraction solution [50% Phenol; 50% Extraction buffer (100 mM Tris-HCl pH 8.0; 100 mM NaCl; 20 mM EDTA; 1% SDS] at 65°C for 10‘ and centrifuged at 14,000 rpm for 10’. The aqueous phase was transferred to new tubes and extracted twice with phenol:chloroform (1:1) and once with chloroform. After precipitation with ethanol, samples were resuspended in H20. LiCl (2M) precipitation (2M LiCl) was employed to separate RNA (pellet) from genomic DNA (supernatant). Genomic DNA was precipitated with ethanol and resuspended in H20.
GEO Accession
GSM1368809