The SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Pico v2) is designed for efficient preparation of Illumina sequencing libraries from picogram inputs (250 pg–10 ng) of total RNA. Technologies included in this product enable processing of high-quality, partially degraded, or low-quality RNA, including RNA prepared from FFPE or LCM samples. The kit features 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. This updated version of our original SMARTer kit for picogram inputs includes refinements that provide improved sequencing performance on all Illumina platforms and a more user-friendly library-purification workflow.
Ribosomal RNA (rRNA) comprises a significant proportion (~90%) of total RNA samples. Depleting these abundant transcripts from total RNA prior to library construction provides benefits by lowering sequencing costs and improving mapping statistics. However, with very low input amounts, rRNA depletion from total RNA leaves an insufficient amount of material for the preparation of a high-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.
Libraries produced with this kit are configured differently than libraries generated with the previous version, such that they achieve a high percentage of clusters passing filter (%PF) without significant additions of PhiX, even on Illumina platforms using two-channel SBS technology such as NextSeq® and MiniSeq™, and on HiSeq® 3000/4000. This in turn reduces sequencing costs by yielding more biologically relevant sequencing reads per run. Data from the Pico v2 kit also tends to identify more unique transcripts with fewer reads mapping to rRNA and mtRNA and lower duplicate rates. Furthermore, the Pico v2 kit includes a new PCR buffer formulation that increases the efficiency of library purification.
This kit is available in 12, 48, 96, and 192-reaction sizes. High-throughput versions of the kit (Cat. # 634413, 634414) include 12 different reverse primers and 8 different forward primers, allowing library construction with up to 96 different combinations of Illumina indexes. The entire protocol, including library prep and purification, takes only ~6 hours.
Please note that we will continue to provide and support the original SMARTer Stranded Total RNA-Seq Kit - Pico Input Mammalian for your convenience.
Alternate kits for other input amounts: