title: GSM4143189: 295_619_1_2; Homo sapiens; RNA-Seq
library_construction_protocol: Resected excess tumor tissue samples were coded and used for RNA sequencing and glioma stem cells isolation in accordance with the Approval by the Institutional Review Board of UMM. Tumor tissue samples were processed within 40 min after resection and divided into three portions under sterile hood. One portion was snap-frozen in liquid nitrogen and kept at -80°C for RNA sequencing after evaluation by neuropathologist. Second portion was used for histopathological assessments. Third portion was used for isolating glioma stem cells. Frozen tumor samples were routinely homogenized using the Minilys personal homogenizer and the Precellys Evolution homogenizer (Bertin Instruments). Grinding was performed using a combination from 1.4 mm and 2.8 mm ceramic beads and 20 mg of tumor tissue at 6500 rpm for 30s x 3 and 30s x 2 pause (4°C). RNA extraction was carried out using the Precellys Tissue RNA Kit Safety-Line (Peqlab) following the manufacturer's protocol. For library preparations, we used the TruSeq® Stranded Total RNA Library Prep (Illumina®) and the NEBNext® Ultra II Directional RNA Library Prep Kit (New England BioLabs®) according to the manufacturer's recommendations. Different dual index adaptors were used for multiplexing samples in one sequencing run. Library concentrations and quality were measured using Qubit dsDNA high-sensitivity (HS) kit and QIAxcel capillary electrophoresis system with QIAxcel ScreenGel software (QIAGEN).
attributes.tag: GEO Accession
attributes.value: GSM4143189