|This protocol has been validated for library construction from 100 ng – 1 µg total RNA, in 10 µL of RNase-free water.
Three fragmented frozen lung tissue samples were randomly selected from each group for the RNA isolation in 3 replicates. Total RNAs were extracted using TRIzol™ Reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions and random primers were used to generate complementary DNA. After quantitative analysis and quality inspection, KAPA-Stranded RNA-Seq Library Prep Kit (Illumina) was used to construct sequencing libraries. Sequencing was carried out using an Illumina HiSeq 4000 Sequencing System for 150 cycles. After the data preprocessing, gene level fragments per kilobase of exon per million fragments mapped and significant changes in gene and transcript expression were then calculated using Ballgown software. Full paper
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Download the product protocol from Roche Lifesciences for KAPA Stranded RNA-Seq Kits with RiboErase (HMR) below.
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