GSM3145776: KD_PMA; Mus musculus; RNA-Seq
Total RNA was prepared using TRIzol reagent (Invitrogen) according to manufacturer's instructions. The quality of the RNA was assessed by a Agilent 2100 BioAnalyzer. Then mRNA was enriched using Oligo dT magnetic beads for library preparation.  Libraries were prepared for 100bp paired-end sequencing using TruSeq RNA Library Preparation kit (Illumina, CA, USA) according to manufacturer's instructions. mRNA molecules were purified and fragmented from 2μg of total RNA using oligo (dT) magnetic beads. The fragmented mRNAs were synthesized as single-stranded cDNAs through random hexamer priming. By applying this as a template for second strand synthesis, double-stranded cDNA was prepared. After sequential process of end repair, A-tailing and adapter ligation, cDNA libraries were amplified with PCR (Polymerase Chain Reaction). Quality of these cDNA libraries was evaluated with the Agilent 2100 BioAnalyzer (Agilent, CA, USA). They were quantified with the KAPA library quantification kit (Kapa Biosystems, MA, USA) according to the manufacturer's library quantification protocol. Following cluster amplification of denatured templates, sequencing was progressed as paired-end (2×100bp) using Illumina HiSeq2500 (Illumina, CA, USA).
GEO Accession
GSM3145776