cDNA synthesis kits—SMART technology
SMART (Switching Mechanism at 5' End of RNA Template) is a unique technology that allows the efficient incorporation of known sequences at both ends of cDNA during first-strand synthesis, without adapter ligation. The presence of these known sequences is crucial for a number of downstream applications, including amplification, RACE, and library construction. While a wide variety of technologies can be employed to take advantage of these known sequences, the simplicity and efficiency of SMART technology permits unparalleled sensitivity and ensures that full-length cDNA is generated and amplified.
SMART (Switching Mechanism at 5' End of RNA Template) is a unique technology that allows the efficient incorporation of known sequences at both ends of cDNA during first-strand synthesis, without adapter ligation. The presence of these known sequences is crucial for a number of downstream applications, including amplification, RACE, and library construction. While a wide variety of technologies can be employed to take advantage of these known sequences, the simplicity and efficiency of SMART technology permits unparalleled sensitivity and ensures that full-length cDNA is generated and amplified.
Minimal RNA sample handling
The entire SMART protocol is performed by one enzymatic reaction, in a single tube. Your precious RNA is subjected to the least possible handling, minimizing potential degradation risks. The protocol is user-friendly and straightforward with no adaptor ligation, no tailing, and no intervening purification steps.
Full-length cDNA enrichment
Because cDNA synthesis is susceptible to interruption by secondary structures in the template RNA, the 5' ends of genes are typically underrepresented in cDNA synthesized by conventional methods. During SMART cDNA synthesis, truncated products resulting from premature termination of the reverse transcription reaction do not incorporate the SMARTer oligonucleotide and consequently are not amplified during PCR. Thus, cDNA created using our SMART technology and amplified by long-distance PCR is enriched for full-length cDNA. Furthermore, because the 5' SMARTer sequence and modified oligo(dT) primer are not added onto genomic DNA or cDNA transcribed from ribosomal RNA, cDNA that is generated using SMART technology is free of sequences derived from these contaminating agents.
Extraordinary sensitivity and high efficiency
One of the greatest advantages of SMART technology is its increased efficiency compared to traditional technologies such as adapter ligation. Its high efficiency and sensitivity enables you to use a very limited quantity of starting material, such as microdissected tissues, laser-captured cells, biopsy samples, etc. As little as 1–2 ng of total RNA is sufficient for generating a highly representative cDNA pool for different downstream applications.
Single-step procedure
During first-strand SMARTer cDNA synthesis, known universal primer sequences are incorporated at both ends of the cDNA. Following first-strand synthesis, SMARTer cDNA is immediately available for PCR amplification.
Overview
- Start with as little as 1–2 ng of total RNA
- Specific enrichment for full-length cDNA
- Optimized protocol for retaining true gene representation
- Simplified protocol without DNase treatment or a separate adapter-ligation step
More Information
Applications
- Cloning and library construction
- Probe generation
- Subtractive hybridization
- Next-generation sequencing
- Virtual northerns
- Template generation for PCR/qPCR
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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