The SMARTer Stranded Total RNA-Seq Kit - Pico Input Mammalian is designed for efficient preparation of Illumina® sequencing libraries from picogram input amounts (250 pg–10 ng) of total RNA. The technologies in this product are designed to process high-quality, partially degraded, and low-quality (e.g., prepared from FFPE or LCM samples) RNA. The kit has a streamlined workflow that retains strand information and eliminates downstream library prep by incorporating indexes and adapters during the reverse-transcription and PCR-amplification steps.
Ribosomal RNA (rRNA) comprises a significant proportion (~90%) of total RNA samples. Depleting these abundant transcripts from total RNA prior to generating libraries provides benefits by lowering sequencing costs and improving mapping statistics. However, with very low input amounts, initial rRNA depletion from total RNA leaves an insufficient amount of material for the preparation of a good-quality library. The workflow used in this kit incorporates a proprietary technology that depletes ribosomal cDNA (cDNA fragments originating from rRNA molecules) using probes specific to mammalian rRNA and some mitochondrial RNA.
The high-throughput version of the kit (Cat. # 635007) contains 12 reverse primers and 8 forward primers, allowing library construction with up to 96 separate indexes. The specific sequences of the primers depend on the reaction size (version) of the kit you choose. The entire protocol, including library prep and purification, takes only ~5 hours.
Alternate kits for other input amounts: