SENSE mRNA-Seq Library Prep Kit V2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Libraries were created from high quality RNA (RNA integrity number of 7 or greater) using the SENSE mRNA-Seq Library Prep Kit from Lexogen (Lexogen, Vienna, Austria) according to the manufacturer’s protocol. This protocol includes an integrated poly(A) mRNA selection process, and the constructed library maintains the strand specificity. One single RNA-Seq library was constructed for each RNA sample. Library production is initiated by the random hybridization of Starter/Stopper heterodimers to the RNA template, followed by first and second strand cDNA synthesis.

Protocol tips

Libraries were created from high quality RNA (RNA integrity number of 7 or greater) using the SENSE mRNA-Seq Library Prep Kit from Lexogen (Lexogen, Vienna, Austria) according to the manufacturer’s protocol. This protocol includes an integrated poly(A) mRNA selection process, and the constructed library maintains the strand specificity. One single RNA-Seq library was constructed for each RNA sample. Library production is initiated by the random hybridization of Starter/Stopper heterodimers to the RNA template, followed by first and second strand cDNA synthesis.

Publication protocol

RNA-sequencing was performed from hippocampal RNA ( = 4/strain/treatment group). RNA quality was evaluated with an Agilent 2100 Bioanalyzer (Agilent Technologies). Libraries were created from high quality RNA (RNA integrity number of 7 or greater) using the SENSE mRNA-Seq Library Prep Kit from Lexogen (Lexogen, Vienna, Austria) according to the manufacturer’s protocol. This protocol includes an integrated poly(A) mRNA selection process, and the constructed library maintains the strand specificity. One single RNA-Seq library was constructed for each RNA sample. Library production is initiated by the random hybridization of Starter/Stopper heterodimers to the RNA template, followed by first and second strand cDNA synthesis. The cDNA was subject to end repair, adenylation of 3′ ends and adapter ligation. The adapters contained a unique “barcode” for each sample, which allows for multiplexing. The cDNA library quality and size distribution were checked using an Agilent Bioanalyzer and DNA 1000 chip. Library fragment sizes were between 200 and 500 bp, with a peak at B260 bp. All libraries were quantified with a Qubit 2.0 Fluorometer (Life Technologies) and stored at −20°C.

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Manufacturer protocol

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