SMART-Seq Total RNA Pico Input with UMIs (ZapR Mammalian)
NOTE: SMART-Seq Total RNA Pico Input with UMIs (ZapR Mammalian) is a replacement for the SMARTer Stranded Total RNA-Seq Kit v3 - 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 with UMIs (ZapR Mammalian) is designed for efficient preparation of Illumina sequencing libraries from picogram input amounts of high-quality, partially degraded, or low-quality total RNA, including total RNA prepared from FFPE or LCM samples. The kit features a streamlined workflow that retains strand information and incorporates indexes and adapters during the reverse-transcription and PCR-amplification steps. The library design featured in this kit adds an 8 nucleotide (nt) unique molecular identifier (UMI) through the reverse-transcription step to mitigate potential PCR bias as well as to provide additional information for transcript quantification, specifically for true variants and rare mutations.
NOTE: SMART-Seq Total RNA Pico Input with UMIs (ZapR Mammalian) is a replacement for the SMARTer Stranded Total RNA-Seq Kit v3 - 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 with UMIs (ZapR Mammalian) is designed for efficient preparation of Illumina sequencing libraries from picogram input amounts of high-quality, partially degraded, or low-quality total RNA, including total RNA prepared from FFPE or LCM samples. The kit features a streamlined workflow that retains strand information and incorporates indexes and adapters during the reverse transcription and PCR amplification steps. The library design featured in this kit adds an 8 nucleotide (nt) unique molecular identifier (UMI) through the reverse-transcription step to mitigate potential PCR bias as well as to provide additional information for transcript quantification, specifically for true variants and rare mutations.
This kit incorporates our proprietary SMART (Switching Mechanism at the 5’ end of RNA Template) technology and includes refinements to the SMARTer method for stranded RNA-seq that simplify the library preparation workflow and improve sequencing performance. This method was developed to work with either high- or low-quality total RNA, does not require additional rRNA removal methods or kits, and produces sequencing libraries that retain strand-of-origin information. The integrated removal of cDNAs derived from rRNA—typically present in high abundance following cDNA synthesis from total RNA inputs—makes the workflow extremely sensitive, yielding data that is highly reproducible with low mapping to rRNA.
The entire protocol, including library prep and purification, takes only ~6.5 hours.
For your convenience, we have outlined the differences between our low-input total-RNA seq kits below:
- SMART-Seq Total RNA Pico Input with UMIs (ZapR Mammalian) supports an expanded input range of 250 pg–1 µg total RNA.
- SMART-Seq Total RNA Pico Input (ZapR Mammalian) supports the same expanded input range of 250 pg–1 µg total RNA but does not include the use of 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
- Seamless integration of UMIs—accurately identify true variants and rare mutations and mitigate potential PCR amplification bias and sequencing errors
- Versatile—generate reproducible data from RNA of any quality (including FFPE and LCM samples and cell-free RNA)
- Fast, streamlined protocol—go from start to finish in ~6.5 hours and save time during library purification with an improved buffer formulation
- Compatible 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 with UMIs (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
- RNA-seq on all Illumina platforms
- 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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