title: GSM4683712: Tg_D15 P10253_236; Mus musculus; RNA-Seq
library_construction_protocol: The singe cell suspension was labeled with magnetic anti-CD11b (130-049-601) and anti-Glast (130-095-825) beads at dilution suggested by manufacturer and incubated and separated using LS columns also according to manufacturer´s exact instructions. The negative fraction was stained with anti-O4 beads (130-094-543) and isolated using MS columns. Beads, reagents and columns were provided by Milteny Biotec, Bergisch Gladbach, Germany. RNA was isolated using RNeasy Micro Kit (Qiagen, Venlo, Netherlands) according to manufacturer´s instructions with minor modifications to the protocol. In brief, isolated cells were lyzed in 500mL Qiazol following sorting, 100mL chloroform was added and samples spun at 2500 x g for 15min at +4°C in a MaXtract column (Qiagen, Venlo, Netherlands). Mixed with 500mL 70% EtOH and spun at 3000 x g for 30s at +4°C in a MiniElute column, washed with 350mL RW1, on-column digestion for 15 min at room temperature, washed with 350mL RW1, 500mL RLT, followed by 500mL 80% EtOH and spun for 2min. RNA was eluted using 10mM Tris pH7.5. Buffer. RNA libraries were prepared for sequencing using the SMARTer Stranded Total RNA-seq Kit - Pico Input Mammalian (Clontech).
attributes.tag: GEO Accession
attributes.value: GSM4683712