Takara SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian

### Strand-specific Illumina sequencing libraries from picogram RNA inputs of any quality

The SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian is designed for efficient preparation of Illumina sequencing libraries from picogram input amounts (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.

Strand-specific Illumina sequencing libraries from picogram RNA inputs of any quality

The SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian is designed for efficient preparation of Illumina sequencing libraries from picogram input amounts (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 includes 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. The entire protocol, including library prep and purification, takes only about 6 hours.

Ribosomal RNA (rRNA) comprises a significant proportion (as much as 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. Nos. 634413 & 634414) include 12 different reverse primers and 8 different forward primers, allowing library construction with up to 96 different combinations of Illumina indexes.

To allow additional customization and flexibility, we also offer the 96- and 192-reaction sizes of this kit without indexing primers (Cat. Nos. 634418 & 634419). These kits can be used with our SMARTer RNA Unique Dual Index Kits (Cat. Nos. 634451, 634452 & 634457), which are sold separately.

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 and options:

• For 10–100 ng of total RNA, we recommend the SMARTer Stranded Total RNA Sample Prep Kit - Low Input Mammalian
• For 100 ng–1 µg of total RNA, we recommend the SMARTer Stranded Total RNA Sample Prep Kit - HI Mammalian

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Overview

• Accommodates picogram inputs of starting material—Start from 250 pg–10 ng of total RNA from human, mouse, or rat.
• Versatile—Generate reproducible data from RNA of any quality (including FFPE and LCM samples and cell-free RNA).
• Improved sequencing performance—Achieve a high percentage of clusters passing filter (%PF) without adding PhiX and identify more transcripts with fewer duplicates.
• Strand information—Identify each transcript’s strand of origin with very high accuracy.
• Fast, streamlined protocol—Go from start to finish in ~6 hours and save time during library purification with an improved buffer formulation.
• Seamless integration with Illumina sequencing—Generate Illumina-ready libraries with up to 192 combinations of indexes.

Interested in more data and FAQs about this product? Visit the NGS Learning Center.

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Applications

• Robust NGS library construction that retains strand information
• Use for RNA-seq on all Illumina platforms
• Captures coding and noncoding information from total mammalian RNA of any quality, including RNA obtained from FFPE samples

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.