RNA sequencing Human - HEK293T

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

TruSeq Small RNA Library Preparation Kit

Illumina

Upstream tips	Protocol tips	Downstream tips
The purified cDNA library was used for cluster generation on Illumina's Cluster Station and then sequenced on an Illumina GAIIx following the vendor's instructions.	A mature miRNA library was generated using a Truseq small-RNA preparation kit (Illumina, USA) according to Illumina's sample preparation guide.

Upstream tips
The purified cDNA library was used for cluster generation on Illumina's Cluster Station and then sequenced on an Illumina GAIIx following the vendor's instructions.
Protocol tips
A mature miRNA library was generated using a Truseq small-RNA preparation kit (Illumina, USA) according to Illumina's sample preparation guide.

Publication protocol 1 Relevant paper

NEBNext® Multiplex Small RNA Library Prep Set for Illumina®

New England BioLabs

Upstream tips	Protocol tips	Downstream tips
	Starting Material: 100 ng–1 µg Total RNA. Small RNA fragments should have a 5´ phosphate and 3´ OH to ligate and must be free of ATP.

Protocol tips
Starting Material: 100 ng–1 µg Total RNA. Small RNA fragments should have a 5´ phosphate and 3´ OH to ligate and must be free of ATP.

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

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