Rapid Amplification of cDNA Ends (RACE) is a technique used to obtain the full-length sequence of an RNA transcript. RACE can provide the sequence of an RNA transcript from a small 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. The 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 use. 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.
- Easy to Use
The SMARTer RACE protocol is performed in a single tube, and minimizes handling of both your RNA sample and your synthesized cDNA. Total hands-on time is only four hours.
- Complete Kit
The SMARTer RACE 5’/3’ Kit contains all the necessary components (save gene-specific primers) to streamline your process. With this single kit, you can begin first-strand cDNA synthesis and proceed through cloning RACE fragments, recovering successful clones on day two.
- Requires Only 10 ng of Total RNA
Our SMARTer RACE method allows you to utilize small samples, including biopsies, tissue dissections, needle aspirations, and embryonic and rare disease tissues. This optimized protocol significantly reduces non-specific background, and such a reduction is essential when handling very small sample sizes.
- Specific Enrichment for 5' Ends
Our carefully designed, chemically-treated SMARTer Oligo preferentially hybridizes to the 5' ends of the cDNA being synthesized. Using this SMARTer Oligo, our procedure enriches cDNA pools for 5' sequences, thus increasing the likelihood you will amplify the entire sequence of your gene or the upstream regulatory regions.
- No RNA Pretreatment Required
Our SMARTer RACE method requires no RNA pretreatment with DNase. This protocol works with total RNA, as well as with samples that may be contaminated with genomic DNA.
Marathon cDNA Amplification Kit
The Marathon cDNA Amplification Kit method employs a specially-designed adaptor that significantly reduces background and permits both 5'- and 3'-RACE reactions (1, 2) 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. No. 639206).
Clontech also offers 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.