title: GSM8557761: CS13 ventral hind brain biological replicate1; Homo sapiens; RNA-Seq
library_construction_protocol: Samples were collected for two biological replicates on independent occasions . Otic vesicles (left and right) and dorsal hindbrain were isolated from CS13 and CS14 human embryos in cold L-15 medium (Thermofisher, 21083027). Tissues were collected into low-binding tubes with 40ul NEB protection buffer (T2010) on ice and frozen straight way at −80 °C. All the samples were stored at −80 °C before they were sent to Novogene for RNA sequencing. RNA purification, library preparation and RNA Sequencing were carried out at Novogene (www.novogen.com). Generally, tissues were processed using the NEB Monarch kit (T2010) for RNA isolation. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. RNA and cDNA quality were checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina HiSeq 4000, 150bp paired end sequencing Bulk RNA seq
identifiers.namespace: GEO
identifiers.id: GSM8557761_r1