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cDNA Synthesis: SMART™ Technology Overview
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.
Single-step, single-tube protocol for 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. The resulting single-stranded cDNA can then be amplified by long-distance PCR.
The SMART Single-Step Procedure
In all SMART and SMARTer kits, first-strand cDNA synthesis is primed by a modified oligo(dT) primer that contains additional sequence at the 3’ end. When SMARTScribe MMLV RT reaches the 5' end of the mRNA, the enzyme’s terminal transferase activity attaches several additional nucleotides onto the newly synthesized strand of cDNA. Then, the SMART oligo pairs with the extended first strand cDNA tail, and serves as a second template for the RT enzyme to switch to. Following first strand synthesis, SMART technology results in the addition of known sequence at both the 3’ and 5’ ends of the cDNA, producing first-strand cDNA that is immediately available for PCR amplification, RACE, library construction and other molecular biology applications.
SMART enriches for high quality, full-length cDNA
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. Since the terminal transferase activity (and subsequent SMART switching process) occurs preferentially at the 5' ends of eukaryotic mRNAs, truncated products resulting from premature termination of the reverse transcription reaction generally 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. 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.
SMART technology is extraordinarily sensitive, and highly efficient
One of the greatest advantages of SMART technology is its increased addition 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.
Figure 1. SMART(er) cDNA synthesis compared to conventional cDNA synthesis. Unlike conventional cDNA synthesis methods,which involve a multiple enzyme/multiple step procedure, the SMART(er) cDNA synthesis protocol is performed by one reverse transcription reaction, in a single tube, with no adaptor ligation or intervening purification steps. Following PCR amplification, SMARTer cDNA is immediately available for a variety of downstream applications.
| Product Name | Size |
Catalog Number |
Price | Add to Cart | Notice to Purchaser | Required Products |
| SMART cDNA Library Construction Kit | each | 634901 | $853.00 USD |
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| SMARTer RACE cDNA Amplification Kit | 10 rxns | 634923 | $734.00 USD |
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| SMARTer RACE cDNA Amplification Kit | 20 rxns | 634924 | $1,174.00 USD |
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| SMARTer PCR cDNA Synthesis Kit | 10 rxns | 634925 | $757.00 USD |
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| SMARTer PCR cDNA Synthesis Kit | 20 rxns | 634926 | $1,211.00 USD |
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| SMARTer Pico PCR cDNA Synthesis Kit | 10 rxns | 634928 | $1,180.00 USD |
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| In-Fusion SMARTer cDNA Library Construction Kit | 10 rxns | 634929 | $1,210.00 USD |
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| SMARTer Fluorescent Probe Amplification Kit | 10 rxns | 634930 | $920.00 USD |
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| SMARTer Pico Fluorescent Probe Amplification Kit | 10 rxns | 634931 | $1,280.00 USD |
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| In-Fusion SMARTer Directional cDNA Library Construction Kit | 10 rxns | 634933 | $1,210.00 USD |
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| SMART mRNA Amplification Kit | 10 rxns | 635001 | $650.00 USD |
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| pRetro-Lib Vector | 20 µg | 635002 | $341.00 USD |
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| pEXP-Lib Vector | 20 µg | 635003 | $341.00 USD |
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| RNA/cDNA Quality Assay | 40 rxns | 636841 | $405.00 USD |
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