GSM5370897: ADA PRMT5 Rep2; Homo sapiens; RNA-Seq
Rneasy Plus Mini Kit (Qiagen), according to the manufacturer's instructions Following plating, 2uL of ERCC controls (using a 1:1,000 dilution) were spiked into each sample. An aliquot of 200ng for each sample was transferred for library preparation which uses automated variant of the Illumina TruSeq Stranded mRNA Sample Preparation kit, preserving strand orientation of the RNA transcript. Oligo dT beads are used to select mRNA from total RNA. It is followed by heat fragmentation and cDNA synthesis from the RNA template. The resultant 400bp cDNA then goes through a dual-indexed library preparation: “A” base addition, adapter ligation using P7 adapters, and PCR enrichment using P5 adapters. After enrichment, the libraries were quantified by Quant-iT PicoGreen (1:200 dilution). After normalizing the samples to 5ng/uL, the set was pooled and quantified using a KAPA Library Quantification Kit for Illumina Sequencing Platforms. The above process is carried out in 96well format and automated pipetting is performed by either an Agilent Bravo or Hamilton Scarlet.
GEO Accession
GSM5370897