title: GSM2640772: S13290 left midbrain; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted at HDBR Newcastle. Tissue samples were divided into sub-samples each weighing thirty milligrams (mg). The sub-samples were homogenised using a Precellys 24 bead mill homogeniser (Bertin Corp. Rockville, MD, USA) using ceramic 1.4mm beads for soft tissue homogenising (CK14) with 600ul of RTL plus Buffer with 10µl/ml of β-Mercaptoethanol and 5µl/ml of reagent DX (Qiagen, Venlo, the Netherlands). RNA and DNA was extracted from the tissue with a QIAcube using an AllPrep DNA/RNA Mini Kit (Qiagen) following the manufacturer’s recommended protocol. RNA was then pooled from all 30 mg sub-samples belonging to a given embryo's left or right midbrain, or left or right forebrain.  RNA quality was assessed using an Agilent 2100 Bioanalyzer (Applied Biosystems, Santa Clara, CA, USA), and then RNA was shipped on dry ice to Beijing Genomics Institute (BGI) Shenzhen/HongKong, China. Oligo(T)'s were used to enrich for mRNA out of the pool of total RNA. After fragmentation, cDNA was generated with random hexamer primers, and the cDNA fragments were connected with adapters, all with the usual quality control steps. Barcoded cDNA fragments were sequenced in two lanes of an Illumina HiSeq 4000 sequencer.  We used paired-end sequencing with a read length of 100 bases.
attributes.tag: GEO Accession
attributes.value: GSM2640772