Rapid Amplification of cDNA Ends (RACE) is a technique used to obtain the full-length sequence of an RNA transcript found within a cell. 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.
SMARTer RACE cDNA Amplification Kit
The SMARTer RACE cDNA Amplification Kit is an improved version of the original SMART RACE cDNA Amplification Kit. The SMARTer RACE Kit provides better sensitivity, less background and higher specificity, due to the inclusion of a newly designed SMARTer II A oligonucleotide and SMARTScribe Reverse Transcriptase, allowing you to perform 3’ or 5’ RACE of your target transcript from as little as 10 ng of total RNA. Additionally, the new SMARTer RACE Kit includes random primers.The SMARTer RACE kit provides an elegant method for incorporating synthetic adaptors into both the 5' and 3' ends of cDNA during synthesis. The 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 result is a powerful method for cloning complete cDNAs that offers several advantages:
- Unprecedented Ease of Use
With our SMARTer RACE method, only a single tube is needed to perform the two-step procedure. Only minimal handling of both your RNA sample and the synthesized cDNA is required. Total hands-on time is only four hours.
- 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.
Clontech also offers Marathon-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.