GSM2898354: Wt_3_SFs; Mus musculus; RNA-Seq
Ankle joints and aortic heart valves were collected, isolated and cultured for 4 passages from the following genotypes: wt, Tnf ARE/+ and Tnf ARE/+ ;Tnfrsf1b MCKO . 3 mice per genotype were used. All samples were diluted to a mean concentration of approximately 100-150ng/μl and their quality was measured in the bioanalyzer (Agilent Technologies) using the Agilent RNA 6000 Nano Kit reagents and protocol(Agilent Technologies). RNA samples with RNA Integrity Number (RIN) were selected and further processed for library preparation and sequencing. Libraries were prepared with the 3' mRNA-Seq Library Prep Kit Protocol for Ion Torrent (QuantSeq-LEXOGEN ™ Vienna, Austria) according to manufacturer's instructions. Each library's quality and quantity, was assessed in bioanalyzer using the DNA High Sensitivity Kit reagents and protocol (Agilent Technologies). The quantified libraries were pooled together in 12plex at a final concentration of 50pM. The pools were then processed, templated and enriched on an Ion Proton Chef instrument. Templating was performed, using the Ion PI™ IC200™ Chef Kit (ThermoFisher Scientific) and sequencing, with the Ion PI™ Sequencing 200 V3 Kit on Ion Proton PI™ V2 chips (ThermoFisher Scientific) according to commercially available protocols. 3'RNA-sequencing was performed on an Ion Proton™ System (Rothberg et al, 2011), according to the manufacturer's instructions.
GEO Accession
GSM2898354