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

Typically 200–300 ng amplified cDNAs were sonicated to around 200 bp long fragments using a Covaris acoustic shearing instrument and then used to construct Illumina sequencing libraries following Illumina’s TruSeq DNA sample preparation protocols. The fragmented cDNAs were end-repaired, a single A base was added to the 3′ end, and then ligated with illumina PE adaptors. Then the ligated DNA fragments were enriched and amplified with ten cycles of PCR. The libraries were sequenced on either Illumina’s HiSeq 2000, or HiSeq 2500 instruments with 100-bp pair-end reads. All clusters that passed the quality filter were exported into fastq files.

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Manufacturer protocol

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

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