Rapid amplification of cDNA ends (RACE) is a technique used to obtain the full-length sequences of RNA transcripts. RACE can provide the sequence of an RNA transcript from a short, known sequence within the transcript all the way to the 5" end (5" RACE-PCR) or 3" end (3" RACE-PCR) of the RNA.
The SMARTer RACE 5"/3" Kit is an improved version of the SMARTer RACE cDNA Amplification Kit. The SMARTer RACE 5"/3" Kit provides improved sensitivity, less background and higher specificity over the previous generation of kits. First-strand cDNA generated with this kit can be used directly in 5"- and 3"-RACE PCR reactions. The SMARTer protocol does not involve any of the adaptor ligation steps that other RACE kits incorporate, making the protocol shorter and significantly easier to execute. The new kit is also designed to accommodate larger RNA input volumes (up to 11 µl) and has been shown to perform more efficiently on challenging targets (e.g., those that are long, GC-rich, etc.).
Rapid amplification of cDNA ends (RACE) is a technique used to obtain the full-length sequences of RNA transcripts. RACE can provide the sequence of an RNA transcript from a short, known sequence within the transcript all the way to the 5" end (5" RACE-PCR) or 3" end (3" RACE-PCR) of the RNA.
The SMARTer RACE 5"/3" Kit is an improved version of the SMARTer RACE cDNA Amplification Kit. The SMARTer RACE 5"/3" Kit provides improved sensitivity, less background and higher specificity over the previous generation of kits. First-strand cDNA generated with this kit can be used directly in 5"- and 3"-RACE PCR reactions. The SMARTer protocol does not involve any of the adaptor ligation steps that other RACE kits incorporate, making the protocol shorter and significantly easier to execute. The new kit is also designed to accommodate larger RNA input volumes (up to 11 µl) and has been shown to perform more efficiently on challenging targets (e.g., those that are long, GC-rich, etc.). The kit includes our highly robust SeqAmp DNA Polymerase for amplification of products, and the In-Fusion HD Cloning Kit for efficient cloning of RACE fragments in the provided linearized vector. The SMARTer RACE 5"/3" procedure combines SMART first-strand cDNA synthesis technology with a powerful suppression PCR protocol that vastly reduces the background amplification commonly associated with RACE protocols.
The Marathon cDNA Amplification Kit method employs a specially designed adaptor that significantly reduces background and permits both 5"- and 3"-RACE reactions ((Bertling, Beier, and Reichenberger 1993; Frohman 1993) to be performed using the same template. Marathon cDNA amplification can be used to quickly characterize multiple RNAs identified by expressed sequence tags (ESTs), differential display, RNA fingerprinting, or cDNA subtraction. Marathon cDNA synthesis begins with poly A+ RNA and a modified lock-docking oligo(dT) primer that contains two degenerate nucleotides at the 3" end. These nucleotides position the primer at the beginning of the poly A tail, eliminating the 3" heterogeneity inherent with conventional oligo(dT) priming. Following cDNA synthesis, blunt ends are created and the Marathon Adaptor is ligated to both ends of the double-stranded cDNA.
NOTE: The Marathon cDNA Amplification Kit requires the Advantage 2 PCR Kit (Cat. # 639206).
We also offer RACE-ready cDNAs, which are double stranded cDNAs made from high-quality premium poly A+ RNA and ligated to the Marathon Adaptor. These cDNAs are ready for 5"- and 3"-RACE PCR and are available from a wide range of tissues and cell types.
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