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.
- 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, which is helpful for analysis of small inputs.
- 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.
Related products
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 ((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).
RACE-ready cDNAs
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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采用抗-HBe抗体包被反应板，加入校准品及被测样本，同时加入定量HBeAg中和抗原，经过振荡孵育,洗板后再加入铕标记的抗-HBe，若标本中抗-HBe浓度高，HBeAg将被大量中和，使最后形成的抗-HBe-HBeAg-铕标记抗-HBe复合物减少。增强液（β－NTA）将标记在抗体上的Eu3+解离到溶液中，Eu3+和增强液中的有效成分形成高荧光强度的螯合物，荧光强度和样本中的抗-HBe浓度成反比。
GM实验：试剂说明书上要求20分钟显色，10分钟的时候看一下，差不多就可以加终止液，但是我的标曲显色特别快，2分钟最低浓度蓝色就就深了，此时样本颜色还很浅很浅，我只能一块儿加终止液。所以，标曲显色过快，可能是什么原因？
（ps:第一列是标曲，后面是样本）
