SMART technology overview
What is SMART technology?
SMART (Switching Mechanism at the 5′ end of RNA Template) technology leverages the template-switching capability of certain reverse transcriptases (RTs) to capture full-length sequence information from RNA or DNA templates and incorporate adapter sequences during first-strand cDNA synthesis. The adapter sequences serve as primer-binding sites during subsequent rounds of PCR amplification, allowing for seamless, ligation-free NGS library construction. SMART technology has become an industry-leading solution for diverse NGS applications by virtue of its versatility and best-in-class performance. See below for more detailed information about how SMART technology works.
Why adopt SMART technology?
- Exquisite sensitivity and reproducibility
- Full-length template coverage
- Ligation-free adapter incorporation
- Strand-of-origin information
- High-quality sequencing libraries
- Simplified workflows, single-tube protocols
Select a topic to learn more:
SMART technology workflow
SMART technology workflow for single-cell/ultra-low-input RNA-seq. The above diagram depicts the workflow for the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing, which uses SMART technology and oligo(dT) priming to generate cDNA from single cells or ultra-low inputs of total RNA. During first-strand cDNA synthesis, the MMLV-derived reverse transcriptase (RT) adds non-templated nucleotides (depicted by Xs) upon reaching the 5' end of the RNA template. A carefully designed template-switching oligonucleotide—in this case, the SMART-Seq v4 Oligonucleotide—then pairs with these additional nucleotides, providing a new template for the RT to add an adapter sequence to the 3' end of the first-strand cDNA (this is the template-switching step). Adapter sequences derived from the template-switching oligonucleotide and oligo(dT) primer then serve as primer-binding sites during subsequent PCR amplification. Other applications of SMART technology involve similar workflows, but use a random-primed mechanism and incorporate Illumina adapter and index sequences during library amplification.
SMART technology can be used for a wide array of NGS applications:
- Single-cell mRNA-seq: solutions for generating sequencing libraries from single cells, small groups of cells, or ultra-low RNA inputs. Integration of oligo(dT) priming and SMART technology ensures full-length, unbiased mRNA coverage, regardless of the presence of rRNA or genomic DNA.
- Total RNA-seq: kits for library prep from total RNA using random priming (with random hexamer oligos) acquire strand-of-origin information through the attachment of SMART adapters during reverse transcription. Illumina-specific sequencing indexes are incorporated during PCR amplification in a ligation-independent manner. This method allows for the identification of both non-coding and coding RNAs.
- Small RNA-seq: SMART technology provides a powerful alternative to adapter-ligation-based methods that are susceptible to sample representation bias. This approach is suitable for analysis of diverse small RNA species, including miRNAs, siRNAs, piRNAs, and snoRNAs.
- Immune profiling: kits feature a 5' RACE-based approach that captures full-length sequence information for entire variable regions of TCR-α and TCR-β subunits. SMART technology provides the sensitivity necessary for detection of low-abundance TCR clonotypes.
- ChIP-seq: kits for generating double-stranded, ready-to-use sequencing libraries from double- or single-stranded chromatin-immunoprecipitated (ChIP) DNA. Illumina-specific sequencing indexes are incorporated during PCR amplification in a ligation-independent manner.
Collaborations & partnerships
Are you interested in custom solutions involving SMART technology?
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