GSM1613346: B6d1_3; Mus musculus; RNA-Seq
20µg of total RNA was enriched for poly A+ RNA using one cycle of the Poly A Purist Kit from Ambion according to manufacturer’s standard protocol. The resulting enriched RNA samples were then analyzed on an Agilent Bioanalyzer to determine   the remaining amount of rRNA in the samples. If the amount was higher than 5%, samples were subjected to another cycle of Poly A enrichment.  100ng of the poly A+ enriched RNA was then used to prepare libraries for sequencing using the AB Library Builder™ Whole Transcriptome Core Kit for 5500 Genetic Analysis Systems on a Library Builder system. Libraries were amplified for 15 cycles before 5500 Wildfire primers were added using five cycles of fusion primer amplification as directed in the 5500 Wildfire manual. Before sequencing, small aliquots of libraries were pooled and sequenced on an Ion Torrent PGM 314 chip after additional amplification with PGM fusion primers. The library pools were quantified by Real-Time PCR and immobilized on flowcells for the SOLiD 5500 Wildfire instrument (Applied Biosystems) and sequenced (50bp reads). The average number of reads per sample was 29.5. One sample had high number of reads (230 million) and the others on average had 23 million reads.
GEO Accession
GSM1613346