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

RNA-seq was performed for MIA PaCa-2 and PANC-1 cells treated with 5-aza-dC or IR exposure (2 Gy and 4 Gy) and both combination. Briefly, total RNA was extracted from MIA PaCa-2 and PANC-1 cell lines with different conditions. For the Illumina library preparations, the extracted RNAs was synthesized using TruSeq RNA sample preparation kit (Illumina Inc., CA, USA) and sequenced using Illumina HiSeq 2000. Paired-end reads were trimmed by quality score using Trimmomatic and mapped to the human reference genome (hg38) using HISAT2. Gene expression levels were quantified by means of the reads per kilo base per million mapped reads (RPKM) method. Differentially expressed genes between different conditions were identified using the GFOLD algorithm. Lowly expressed genes which were below 1 RPKM were discarded.

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Manufacturer protocol

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

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