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Selection Guide: Products for Next-Gen Sequencing

TECHNOLOGY OVERVIEW

SMART Tools for Next-Gen Sequencing

Takara Clontech kits for next-generation sequencing (NGS) offer exceptional sensitivity and reproducibility, and with greater confidence in their data, scientists can spend more time pursuing the questions they care about.

What is SMART technology?

The unparalleled sensitivity of SMARTer NGS kits is achieved through the application of SMART (Switching Mechanism at the 5′ end of RNA Template) technology. SMART technology leverages the template-switching capability of a reverse transcriptase (RT) to capture full-length RNA transcripts and incorporate adapter sequences during first-strand cDNA synthesis, such that sequencing libraries can be generated seamlessly via subsequent rounds of PCR amplification. The best-in-class performance of SMART technology has positioned it as a leading solution for ultra-low input RNA-seq.

Why adopt SMART technology?

  • Enjoy high sensitivity and reproducibility
  • Obtain full-length transcript information for isoform and splice-variant detection
  • Prepare high-quality sequencing libraries
  • Acquire strand-of-origin information

Select a topic to learn more

  • During first-strand cDNA synthesis, the MMLV-derived RT adds non-templated nucleotides when it reaches the 5′ end of the RNA template.
  • RNA templates are reverse transcribed in a 5′ cap-independent manner, which allows for comprehensive sample representation and a streamlined workflow.
  • Hybridization between the SMART-Seq v4 Oligonucleotide and the non-templated nucleotides is stabilized by LNA technology, which has further improved the sensitivity of our latest kits.
  • The SMART-Seq v4 Oligonucleotide provides a new template for the RT to add an adapter sequence to the 3′ end of the full-length cDNA (this is the template-switching step).
  • Adapter sequences at both ends of the first-strand cDNA serve as priming sites for PCR amplification.
  • Depending on the experimental approach, PCR amplification yields either cDNA libraries that are ready for the addition of sequencing indexes, or libraries which already include Illumina®-compatible indexes (which have been incorporated in a ligation-free manner).

This schematic represents the SMART method for cDNA synthesis from mRNA, which utilizes an oligo(dT) primer (3' SMART-Seq CDS Primer IIA) for reverse transcription and the SMART-Seq v4 Oligo for template switching. For applications involving capture of total RNA, the core mechanism is the same and a random primer (N6 primer) is used for reverse transcription.

SMART technology can be used for a variety of applications, including single-cell RNA-seq, total RNA-seq, and ChIP-seq.

  • 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.
  • 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.

Are you interested in custom solutions involving SMART technology?

Whether you seek OEM partnership, custom formulation, or bulk purchasing, our entire OEM team stands behind one promise: we will help you achieve your goals. Our dedicated manufacturing, packaging, QA, and business development professionals stand ready to help. Find out how our technologies and logistics can ensure your success: submit an online inquiry today or call us at 650.919.7320.

Partner with Clontech

We are always seeking partnerships and collaborations that will yield high-quality, innovative tools and services for our customers and accelerate scientific progress. If you have a technology, product idea, or project that you would like to discuss, please contact us by calling 650.919.7320 or e-mailing BD_OEM@clontech.com. One of our Business Development professionals will follow up with you shortly. Kindly provide non-confidential information only in your initial inquiry.




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