TruSeq Stranded Total RNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	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

RNA was extracted with TRIzol reagent (Ambion) and purified with the Direct-zol RNA MiniPrep kit (Zymo Research). DNase I treatment was performed on the Direct-zol column. RNA concentration was determined with a Nanodrop apparatus (Thermo Fisher Scientific) and its integrity (size distribution) with a Bioanalyzer 2100 (Agilent).
Libraries were prepared using TrueSeq stranded total RNA sample preparation (Illumina). Sequencing was performed on an Illumina HiSeq 2000 sequencer with 100 bp single-end reads. Reads were aligned with TopHat (v2.0.7) to the mouse genome (v10) or the human genome (v19). 7SL and RNase MRP reads were quantified using Bedtools (v2.15.0).

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Manufacturer protocol

Download the product protocol from Illumina for TruSeq Stranded Total RNA below.

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