Strand-specific RNA-seq libraries for Illumina platforms from high-input total RNA
The SMARTer Stranded Total RNA Sample Prep Kit – HI Mammalian provides a rapid solution for preparing indexed Illumina sequencing libraries from 100 ng–1 µg of mammalian total RNA. This kit combines RiboGone and SMART technologies, seamlessly incorporating efficient rRNA removal followed by strand-specific library generation in around 5 hours. Because rRNA can comprise ~90% or more of total RNA, effective depletion of rRNA prior to library generation can lower sequencing costs and improve mapping statistics. SMART cDNA synthesis technology followed by PCR complete a streamlined process that performs strand information and eliminates downstream library prep by incorporating indexes and adapters during the reverse transcription and amplification steps.
These high-input kits contain 12 reverse primers and up to 8 forward primers that allow for multiplexing of up to 96 separate libraries in a high-throughput format. The specific primers you receive will depend on your particular kit. The entire protocol—from rRNA depletion through creation of Illumina-specific, indexed RNA-seq libraries—takes only about 5 hours.
Overview
- Strand information—Identify each transcript’s strand of origin with >99% accuracy.
- Fast, streamlined protocol—Go from start to finish in ~5 hours.
- Broad input range—Use 100 ng–1 µg of total RNA from human, mouse, or rat.
- Versatility—Get reliable data across replicates. See high-quality, reproducible data from total RNA with a RIN (RNA Integrity Number) from 3–10.
- Seamless integration with Illumina sequencing—Generate up to 192 uniquely indexed Illumina-ready libraries.
Applications
- RNA-seq for mammalian samples on Illumina platforms
- NGS library generation that retains strand information
- Analysis of coding and noncoding sequence information
