SMART-Seq Total RNA Pico Input (ZapR Mammalian)
NOTE: SMART-Seq Total RNA Pico Input (ZapR Mammalian) is a replacement for SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian with with some updates for improved performance as indicated below:
- Improved efficiency of ZapR rRNA depletion reagents for better removal of unwanted reads
- New kit sizes match those of Unique Dual Index kits (24 rxns, 96 rxns, 384 rxns)
- For added flexibility, indexes are not included in the kit. A choice of indexing primers are sold separately (see Unique Dual Index kits)
This kit can now be used for library preparation with higher input amounts—up to 1 µg of total RNA—while maintaining the same exceptional performance. See the alternative protocol below or download our new flyer for the data.
SMART-Seq Total RNA Pico Input (ZapR 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.
NOTE: SMART-Seq Total RNA Pico Input (ZapR Mammalian) is a replacement for SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian with some updates for improved performance as indicated below:
- Improved efficiency of ZapR rRNA depletion reagents for better removal of unwanted reads
- New kit sizes match those of Unique Dual Index kits (24 rxns, 96 rxns, 384 rxns)
- For added flexibility, indexes are not included in the kit. A choice of indexing primers are sold separately (see Unique Dual Index kits)
This kit can now be used for library preparation with higher input amounts—up to 1 µg of total RNA—while maintaining the same exceptional performance. See the alternative protocol below or download our new flyer for the data.
SMART-Seq Total RNA Pico Input (ZapR Mammalian) is designed for efficient preparation of Illumina sequencing libraries from picogram input amounts of total RNA. Technologies included in this product, including our proprietary SMART (Switching Mechanism at the 5′ end of RNA Template) technology, 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. The entire protocol, including library prep and purification, takes only ~6.5 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 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 kit tends to identify more unique transcripts with fewer reads mapping to rRNA and mtRNA and lower duplicate rates.
For your convenience, we have outlined the differences between our low-input total-RNA seq kits below:
- SMART-Seq Total RNA Pico Input (ZapR Mammalian) supports an expanded input range of 250 pg–1 µg total RNA.
- SMART-Seq Total RNA Pico Input with UMIs (ZapR Mammalian) supports the same expanded input range of 250 pg–1 µg total RNA and incorporates UMIs in the library prep.
- SMART-Seq Total RNA Single Cell (ZapR Mammalian) supports an input range of 1–1,000 cells or 10 pg–10 ng total RNA
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 ~6.5 hours and save time during library purification with an improved buffer formulation.
- Seamless integration with Illumina sequencing—generate Illumina-ready libraries with up to 384 combinations of indexes.
More Information
Alternative protocol for expanded input range between 10 ng–1 µg
To prepare libraries from inputs within 10 ng–1 µg, we recommend the following modifications to the number of cycles for PCR 1 (cDNA amplification; Section V.B of the user manual) and PCR 2 (library amplification; Section V.E of the user manual), below. For all other protocol steps, please follow the Smart-Seq Total RNA Pico Input (ZapR Mammalian) User Manual.
| Input | PCR 1 cycles | PCR 2 cycles* |
| >10 ng–100 ng | 5 | 14 |
| >100 ng–200 ng | 5 | 12 |
| >200 ng–1 μg | 3 | 12 |
*PCR 2 cycles may be increased if library yields are low. See the user manual for more information.
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.
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