GSM2045891: 2818-CPP-0019 L635; Mus musculus; RNA-Seq
Stomach corpus was isolated and diced into 1mm cubed pieces, ezymatically digested for 7 minutes in DMEM/Dispase.  Stomach then put over a 70micron filter and pushed through filter using rubber syringe.  Cells then spun down, washed, antibody labeled and FAC sorted. RNA samples that had RNA integrity numbers (RIN) ≥7.0 were used. RNA was amplified using the NuGEN Technologies Ovation RNA-Seq System V2 kit (Part # 7102-08) using manufacturer's instructions.  Approximately, 2.5 µg amplified DNA from each sample was sheared on a Covaris S200 focused-ultrasonicator (Woburn, MA, USA) with a target yield of an average 200bp fragment size. The fragmented DNA was taken into standard library preparation protocol using NEBNext® DNA Library Prep Master Mix Set for Illumina® (New England BioLabs Inc., Ipswich, MA, USA) with slight modifications. Briefly, end-repair was done followed by polyA addition and custom adapter ligation. Post-ligated materials were individually barcoded with unique in-house genomics service lab (GSL) primers and amplified through 8 cycles of PCR using KAPA HiFi HotStart Ready Mix (Kapa  Biosystems,  Inc.,  Woburn,  MA,  USA).  The quantity of the libraries were assessed by Qubit® 2.0 Fluorometer, and the quality of the libraries was estimated by utilizing a DNA 1000 chip on an Agilent 2100 Bioanalyzer, respectively. Accurate quantification for sequencing applications was determined using the qPCR-based KAPA Biosystems Library Quantification kit (Kapa Biosystems, Inc., Woburn, MA, USA). Each library was then diluted to a final concentration of 12.5nM and pooled equimolar prior to clustering. Single End (SE) sequencing was performed on an Illumina HiSeq2500 following the manufacturer's protocols.
GEO Accession
GSM2045891