RNA sequencing Rat - PC12

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.

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Found 3 matching solutions for this experiment

Ion Total RNA-Seq Kit v2

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	100 ng–50 μg total RNA, we recommend using the DynabeadsTM mRNA DIRECTTM Micro Kit Micro Kit (Cat. No. 61021). 50–400 μg total RNA, we recommend performing two rounds of oligo(dT) selection of the poly(A) RNA using the MicroPoly(A)PuristTM Kit (Cat. No. AM1919). Also, confirm the absence of 18S and 28S rRNA; for example, check the profile of the poly(A) RNA on an AgilentTM 2100 BioanalyzerTM instrument.

Protocol tips
100 ng–50 μg total RNA, we recommend using the DynabeadsTM mRNA DIRECTTM Micro Kit Micro Kit (Cat. No. 61021). 50–400 μg total RNA, we recommend performing two rounds of oligo(dT) selection of the poly(A) RNA using the MicroPoly(A)PuristTM Kit (Cat. No. AM1919). Also, confirm the absence of 18S and 28S rRNA; for example, check the profile of the poly(A) RNA on an AgilentTM 2100 BioanalyzerTM instrument.

Protocol tips

100 ng–50 μg total RNA, we recommend using the DynabeadsTM mRNA DIRECTTM Micro Kit Micro Kit (Cat. No. 61021).
50–400 μg total RNA, we recommend performing two rounds of oligo(dT) selection of the poly(A) RNA using the MicroPoly(A)PuristTM Kit (Cat. No. AM1919). Also, confirm the absence of 18S and 28S rRNA; for example, check the profile of the poly(A) RNA on an AgilentTM 2100 BioanalyzerTM instrument.

Manufacturer protocol Publication protocol 1 Relevant paper

TruSeq RNA Library Prep Kit v2

Illumina

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)

Manufacturer protocol Publication protocol 1 Relevant paper

NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina®

New England BioLabs

Upstream tips	Protocol tips	Downstream tips
	Starting Material: 100 ng-1μl total RNA in a 12 μl total volume.

Protocol tips
Starting Material: 100 ng-1μl total RNA in a 12 μl total volume.

Manufacturer protocol Publication protocol 1 Relevant paper

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