TruSeq RNA Library Prep Kit v2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	0.1 - 1 ug total RNA or 10 - 400 ng previously isolated mRNA (from species with polyA tails)

Protocol tips
0.1 - 1 ug total RNA or 10 - 400 ng previously isolated mRNA (from species with polyA tails)

Publication protocol

Total RNA was extracted with TRIzol reagent following the recommendations of the manufacturer. The quality of total RNA was evaluated using the Agilent 2100 bioanalyzer (Agilent, Palo Alto, CA, USA) with the RNA 6000 Nano LabChip kit. RNA-seq libraries were prepared Illumina® TruSeq RNA Library Prep Kit v2 and were sequenced using Illumina® Hiseq2500 to obtain 150-bp paired-end reads. The sequencing depth for each sample was >20 million reads. The reads were aligned with TopHat 2.0.13 to GRCh37 with default parameters, and then were assembled by Cufflink 2.2.1, using Ensembl v75 annotations. Transcript abundance was measured in fragments per kb of exon per million fragments mapped (FPKM). The RNA-seq data is available at GEO (GSE100687).

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Manufacturer protocol

Download the product protocol from Illumina for TruSeq RNA Library Prep Kit v2 below.

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