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

Three sets of NSC34 cells (transfected with wild-type or G93A mutant hSOD1) were grown and harvested with each set from a separate passage of single cell line. Following subcellular fractionation, transcriptomes of 12 samples were analysed by RNA-seq (Macrogen Inc. Seoul, Korea), as described previously. Briefly, 1 μg of the total RNA was analysed using the TruSeq RNA library kit to construct the cDNA libraries. The protocol included polyA-selected RNA extraction, RNA fragmentation, random hexamer primed reverse transcription and 100 nt paired-end sequencing using an Illumina HiSeq2000 (Illumina, San Diego, CA, USA). The libraries were quantified using quantitative real-time polymerase chain reaction (qPCR) according to the qPCR Quantification Protocol Guide. An Agilent Technologies 2100 Bioanalyzer was used for the qualification.

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Manufacturer protocol

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

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