ScriptSeq Complete Kit (Human/Mouse/Rat)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The Ribo-Zero process uses 100 ng - 1 μg of DNA-free RNA.

Protocol tips
The Ribo-Zero process uses 100 ng - 1 μg of DNA-free RNA.

Publication protocol

Approximately 500 ng of RNA from each sample was used to prepare individually barcoded strand-specific RNA-seq libraries. rRNA and mitochondrial RNA was removed using Ribo-Zero Gold Removal Magnetic Kit for human/mouse/rat (Epicentre, now Illumina) following the manufacturer’s instructions. mRNA was purified using Agencourt RNAClean XP kit. Libraries were constructed using ScriptSeq™ Complete Kit (Epicentre, now Illumina) as per manufacturer’s instructions. Quality and quantity of the libraries were determined using a Bioanalyzer (Agilent) and Qubit® (Invitrogen), respectively. Libraries were then quantified by qPCR in an Applied Biosystems® 7500 with the Kapa Biosystems kit for library quantitation (Kapa Biosystems). Next-generation sequencing was performed using an Illumina HiSeq 2000 flow cell with one 150-bp end run. The NCBI Gene Expression Omnibus (GEO) accession number for the RNA-seq data reported in this paper is GSE88801.

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Manufacturer protocol

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