Three-prime Race
Three-prime Race - Related Videos
06:16
Verbal Priming
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2023
Source: Laboratory of Jonathan Flombaum—Johns Hopkins University Human memory seems to work in two broad ways. Like modern computers, the human mind has explicit, or declarative, memory: ask a question, and a person gives the best answer they can. Input a query, and a computer program returns the contents of the relevant parts of its stored memory. Humans also have a second kind of memory system, one not really typical of computers, one that experimental psychologists call implicit. Implicit...
02:35
RACE - Rapid Amplification of cDNA Ends
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2021
Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer. Since the...
13:54
Tractable Mammalian Cell Infections with Protozoan-primed Bacteria
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Cited by 2 •
2013
This technique provides a method to harvest, normalize and quantify intracellular growth of bacterial pathogens that are pre-cultivated in natural protozoan host cells prior to infections of mammalian cells. This method can be modified to accommodate a wide variety of host cells for the priming stage as well as target cell types.
08:13
Using the Race Model Inequality to Quantify Behavioral Multisensory Integration Effects
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Cited by 20 •
2019
The current study aims to provide a step-by-step tutorial for calculating the magnitude of multisensory integration effects in an effort to facilitate the production of translational research studies across diverse clinical populations.
09:34
Intravital Imaging of Neutrophil Priming Using IL-1β Promoter-driven DsRed Reporter Mice
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Cited by 1 •
2016
This current protocol employs fluorescent reporters, in vivo labeling, and intravital imaging techniques to enable monitoring of the dynamic process of neutrophil priming in living animals.
FAQs
The poly(A) tail provides the attachment point for the oligo(dT) portion of the primer during reverse transcription. Because the same primer also carries an adaptor sequence, the resulting cDNA contains a known sequence adjacent to the previously unknown transcript end. This arrangement creates a defined site for subsequent amplification and connects RNA structure to a sequenceable cDNA product.
The gene-specific forward primer selects cDNA derived from the transcript of interest, whereas the adaptor-specific reverse primer recognizes the known sequence introduced during reverse transcription. Their pairing restricts amplification to molecules that contain both the target gene region and the captured 3′ end. This primer design links sequence specificity at one side with universal end recovery at the other.
Sequencing the amplified product can establish the transcript’s 3′ untranslated region, the cleavage site, and the polyadenylation site. These features extend gene-structure information beyond the known internal sequence and help determine where the RNA transcript terminates. The resulting sequence therefore supports a more complete description of transcript architecture and can distinguish transcript forms with different 3′ ends.
Different polyadenylation sites can produce transcript forms with distinct 3′ ends or 3′ untranslated regions. By identifying the endpoint captured from a transcript, Three-prime RACE provides sequence evidence for these alternative arrangements. Comparing the recovered ends can therefore contribute to transcript-diversity analysis and reveal that one gene may be represented by more than one transcript structure.
The workflow begins with polyadenylated RNA and reverse transcription using an oligo(dT) primer linked to an adaptor. The resulting cDNA is then amplified with a gene-specific forward primer and an adaptor-specific reverse primer. Finally, sequencing of the amplified product is used to determine the transcript’s 3′ region and its cleavage and polyadenylation sites.
This approach is particularly useful when researchers know only part of a transcript sequence but need to define its downstream end. It can support transcript annotation, clarify gene structure, and investigate transcript diversity through 3′-end differences. In genetics, those results help connect partial sequence information with a more complete representation of expressed transcript forms.