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 adaptor 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 the single-step SMART process 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 the 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 cDNAs synthesized by conventional methods. During SMART cDNA synthesis truncated products resulting from premature termination of the reverse transcription reaction do not incorporate the SMART(er) 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’ SMART(er) sequence and modified oligo dT primer are not added onto genomic DNA or cDNA transcribed from ribosomal RNA, cDNA that is generated using SMART is free of 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 adaptor 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.
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.