GSM4057116: CC036cd3_3; Mus musculus; RNA-Seq
For tissue preparation, mice were sacrificed and entire lungs were extracted. Lungs were immediately transferred to RNAlater solution, kept at 4°C for one day and subsequently stored at -20°C. RNA was isolated using Qiagen Midi Kit as described previously. RNA quality was controlled on a 2100 Bioanalyzer Instrument (Agilent). 500ng of total RNA was used to prepare libraries for sequencing using the Lexogen SENSE mRNA-seq library kit for Ion Torrent. Libraries amplified for 11 cycles as the final step of library preparation. Before sequencing, 1ul aliquots of this material was pooled and sequenced on an Ion Torrent PGM 314 chip. Barcode quantification data from the PGM were used to balance the barcodes for final pooling before sequencing. Following this final pooling, the library pools were sized to a target size of 260bp on a Pippin Prep instrument using the 2% Pippin Agarose gel. The sized libraries were examined on an Agilent High Sensitivity DNA chip, quantified using real-time PCR, and used to prepare spheres using a One-Touch 2 device. RNA-Seq; These spheres were then sequenced on an Ion Torrent Proton sequencer with a P1 chip. The mean number of reads per sample was 24 million.
GEO Accession
GSM4057116