RNA sequencing Human - H7

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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Protocol tips
Total RNA input must be between 500 pg and 100 ng. Total RNA inputs above
100 ng per reaction may inhibit amplification, while inputs of less than 500 pg will potentially result in insufficient yields depending on the requirements of the
analytical platform.
Downstream tips
RNA samples for use with the Ovation RNA-Seq System V2 must be stored at –80°C.
Avoid frequent freeze/thaw cycles, or RNA degradation may result.
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