3 Prime Race
3 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...
11:05
Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell (MSC) Spheroids Primed in 3-D Cultures Under Xeno-free Conditions
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Cited by 21 •
2017
The therapeutic potential of mesenchymal stem/stromal cells (MSCs) is well-documented, however the best method of preparing the cells for patients remains controversial. Herein, we communicate protocols to efficiently generate and administer therapeutic spherical aggregates or 'spheroids' of MSCs primed under xeno-free conditions for experimental and clinical applications.
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.
12:49
Detection of miRNA Targets in High-throughput Using the 3'LIFE Assay
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Cited by 8 •
2015
Luminescent identification of functional elements in 3’ untranslated regions (3’UTRs) (3’LIFE) is a technique to identify functional regulation in 3’UTRs by miRNAs or other regulatory factors. This protocol utilizes high-throughput methodology such as 96-well transfection and luciferase assays to screen hundreds of putative interactions for functional repression.
FAQs
The oligo(dT) primer anchors reverse transcription at the transcript’s poly(A) tail. This provides a sequence-based route toward the RNA’s 3′ terminus, even when the region beyond the known gene sequence has not been characterized. Its placement allows the resulting cDNA to retain information about the transcript end and supports targeted amplification of that region.
A gene-specific primer binds within the characterized portion of the transcript, while the second primer recognizes the sequence added during reverse transcription. PCR therefore amplifies the segment lying between the known internal sequence and the downstream transcript terminus. Primer placement gives the reaction its target specificity and connects established gene information with the uncharacterized 3′ region.
Results from 3 Prime RACE can help define the 3′ untranslated region, the sequence located after the coding region, and identify alternative polyadenylation sites. They can also reveal differences among transcript isoforms that arise from distinct RNA ends. These observations contribute to reconstructing gene structure and examining how RNA processing affects transcript organization.
By characterizing transcript ends and 3′ untranslated regions, 3 Prime RACE supplies information about RNA features associated with gene regulation. Detecting different polyadenylation sites or transcript isoforms can show that a gene produces more than one processed RNA form. In genetics, these findings help connect RNA processing patterns with differences in gene expression and transcript structure.
The workflow begins by reverse-transcribing RNA with an oligo(dT) primer that binds the poly(A) tail. PCR then uses a gene-specific primer together with a primer complementary to the sequence added during reverse transcription. Amplification targets the interval between the known gene sequence and the transcript end, producing material for characterizing that previously unresolved region.
Researchers would choose this approach when the internal gene sequence is available but the transcript’s downstream end remains uncertain. It is particularly useful for investigating 3′ untranslated regions, alternative polyadenylation, transcript isoforms, or incomplete gene structures. The method extends existing sequence information toward the RNA terminus rather than requiring the entire transcript end to be known beforehand.
The amplified product can help locate the transcript terminus and clarify how the RNA is organized beyond its known sequence. Comparing products can indicate distinct 3′ ends, alternative polyadenylation sites, or different transcript isoforms. Such information supports analysis of gene structure and provides molecular context for RNA processing and expression studies in genetics.