TruSeq Stranded mRNA

Protocol tips

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	The protocol is optimized for 0.1–1 µg of total RNA

Protocol tips
The protocol is optimized for 0.1–1 µg of total RNA

Publication protocol

Gingival samples were collected from two rats in each group from the prevention experiment and processed for RNA sequencing (RNA-Seq) on the Illumina platform. Total RNA was extracted from homogenized gingival tissue with an RNeasy® Mini kit (QIAGEN Inc., Valencia, CA). RNA quality was assessed with the Agilent RNA 6000 NanoLabChip Kit of the Bioanalyzer system (Santa Clara, CA, USA). RNA-Seq cDNA libraries were made using the Illumina Truseq stranded mRNA Sample Prep kit and were sequenced using the NextSeq v3, 2 x 75 cycle cartridge on a NextSeq sequencer. The RNA-Seq reads were uploaded to the Illumina BaseSpace cloud platform (http://basespace.illumina.com) and the RNAExpress application was used to align reads to the rat genome (Rattus norvegicus UCSC version rn5), calculate read count per gene using the STAR aligner, and perform differential gene expression analysis with DESeq2. The raw RNA sequencing reads were also submitted to the National Center for Biotechnology Information Sequence Read Archive database (http://www.ncbi.nlm.nih.gov/Traces/sra/, BioProject number PRJNA321482).

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Manufacturer protocol

Download the product protocol from Illumina for TruSeq Stranded mRNA below.

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