title: GSM4491361: HP424a; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was isolated from biological duplicates when a cumulative at PD~16 for RNA-seq analysis. 106 cells were rinsed twice in PBS and centrifuged at 260 RCF for 10 min. After removing the supernatant, the cell pellet was lysed by adding 350 μl RLT buffer (Qiagen) containing 1% β-mercaptoethanol (Sigma Aldrich) and homogenized by thoroughly pipetting up and down. Samples were directly placed on ice and frozen at -80 °C until further processing. The Qiagen RNeasy mini kit was used to isolate RNA according to the manufacturer's instructions. cDNA was obtained by reverse transcription of 500 ng of total RNA, using the RevertAid H Minus First Strand cDNA Synthesis Kit, 1 mM oligo-dT20 and random hexamer primers (Thermo Fisher Scientific). RNA-seq samples were obtained from 3 donors for the maxillary samples (HP417, HP421 and HP424), 4 donors for mandibular samples (HP418, HP422, HP425 and HP427) and 3 tibial donors (HP415, HP420 and HP423). RNA-seq samples were prepared from hPDCs at cumulative PD~16 in technical duplicates (A and B) and sequencing library was generated using the Illumina TruSeq Stranded mRNA Library Prep Kit. The resulting cDNA libraries were sequenced (50bp length) according to Illumina TruSeq Rapid v2 protocol on an Illumina HiSeq2500 sequencer.
attributes.tag: GEO Accession
attributes.value: GSM4491361