GSM8574731: PHLDA2_1; Homo sapiens; RNA-Seq
Tumors were extracted from mice and digested into a single cell suspension. GFP+ tumor cells were isoalted with FACS. RNA isolation and bulk sequecing preparations were performed on sorted tumor cells. Library construction was performed according to the Illumina TruSeq mRNA stranded protocol.  The input quantity for total RNA within the recommended range and mRNA was enriched using oligo dT magnetic beads. The enriched mRNA was chemically fragmented. First strand synthesis used random primers and reverse transcriptase to make cDNA. After second strand synthesis the ds cDNA was cleaned using AMPure XP beads and the cDNA was end repaired and then the 3' ends were adenylated. Illumina barcoded adapters were ligated on the ends and the adapter ligated fragments were enriched by nine cycles of PCR. The resulting libraries were validated by qPCR and sized by Agilent Bioanalyzer DNA high sensitivity chip. The concentrations for the libraries were normalized and then multiplexed together. The multiplexed libraries were sequenced using paired end 100 cycles chemistry for the Illumina NovaSeq 6000.