GSM2842613: 132_RSS31_JoDo01_HD_D1_F_neg_N721_S506; Mus musculus; RNA-Seq
Total RNA was purified using the Direct-zol RNA MiniPrep Plus and RNA Clean & Concentrator-25 kits (Zymo Research, Irvine, CA). RNA quality and quantity was assessed using an Agilent 2100 Bioanalyzer with a RNA 6000 Pico Kit (Agilent Technologies, Santa Clara, CA) resulting in all samples producing RIN scores above 9.0. 5 ng of total RNA was then amplified using the Smart-seq2 protocol. A 1:1 ratio of Agencourt AMPure XP beads (Beckman Coulter Inc., Brea, CA) were used to purify resulting cDNA, and 1 ng cDNA was subsequently used to prepare a sequencing library with the Nextera XT DNA Library Prep Kit (Illumina, San Diego, CA). Both whole-transcriptome amplification and sequencing library preparations were performed in a 96-well format to reduce assay-to-assay variability. Quality control steps were included to determine total RNA quality and quantity, the optimal number of PCR pre-amplification cycles, and fragment size selection. The resulting libraries were deep sequenced on the HiSeq 2500 System in Rapid Run mode (Illumina) using 50-bp single-end reads (~6.0×10^6 reads per condition).
GEO Accession
GSM2842613