Rna Amplification
Rna Amplification - Related Videos
09:44
Rapid Amplification of cDNA Ends
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2023
Source: Pablo Sanchez Bosch2, Sean Corcoran2 and Katja Brückner1,2,3 1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research 2Department of Cell and Tissue Biology, 3Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Rapid Amplification of cDNA Ends (RACE) is a technique that allows amplification of full-length cDNA from mRNA by extending to the 3’ or 5’ end, even without prior knowledge of the sequence (Frohman et al.,...
03:26
Probe Hybridization and Signal Amplification in RNA In Situ Hybridization: A Technique for Detecting Specific RNA Sequences in Tissue Sections
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2023
This video describes a sequential hybridization strategy that involves hybridizing mRNA probes to specific target mRNA. Following probe hybridization, signal amplification is performed to enhance resolution via reduction of signal-to-noise ratio during RNA-CISH of histological samples.
09:54
RNA-Seq
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2023
Among different methods to evaluate gene expression, the high-throughput sequencing of RNA, or RNA-seq. is particularly attractive, as it can be performed and analyzed without relying on prior available genomic information. During RNA-seq, RNA isolated from samples of interest is used to generate a DNA library, which is then amplified and sequenced. Ultimately, RNA-seq can determine which genes are expressed, the levels of their expression, and the presence of any previously unknown transcripts.
10:12
Protein Misfolding Cyclic Amplification of Prions
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Cited by 7 •
2012
Protein misfolding cyclic amplification (PMCA) is an in vitro assay for the study of prion conversion and strain and species barriers. It can also be used as a prion detection assay.
11:32
Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
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Cited by 20 •
2017
Here, we detail the method of Sequencing of Psoralen crosslinked, Ligated, and Selected Hybrids (SPLASH), which enables genome-wide mapping of intramolecular and intermolecular RNA-RNA interactions in vivo. SPLASH can be applied to study RNA interactomes of organisms including yeast, bacteria and humans.
FAQs
In reverse-transcription workflows, reverse transcriptase uses the RNA template to produce complementary DNA, or cDNA. Polymerase-based amplification then repeatedly copies that cDNA, allowing the original RNA signal to be represented by a much larger product. This two-stage logic separates RNA conversion from subsequent copying and makes downstream detection or sequencing practical when the starting material is scarce.
Some RNA amplification methods avoid the cDNA step by amplifying RNA directly under constant-temperature conditions. This distinguishes them from workflows that first convert RNA and then use polymerase-based copying. The choice matters because the assay design determines which molecular form is amplified and which reaction conditions must remain controlled throughout the process.
Fluorescence, electrophoresis, and sequencing answer different analytical needs after amplification. Fluorescence can support measurement within an assay, electrophoresis can reveal amplified products, and sequencing can provide sequence-level information. Thus, detection is not merely a final technical step: it determines whether the experiment emphasizes quantification, product visualization, or downstream transcript analysis.
A typical RNA amplification workflow starts with a small RNA sample and follows the assay’s chosen conversion route. In a cDNA-based design, reverse transcription precedes polymerase copying; in a direct design, RNA is amplified under constant-temperature conditions. The resulting material is then assessed by fluorescence, electrophoresis, or sequencing, depending on the intended measurement.
RNA amplification is especially valuable when biological samples contain limited or degraded RNA. In gene-expression analysis, it increases the material available for measuring transcripts; in pathogen detection, it can help reveal scarce RNA; and in transcriptome studies, it supports analysis from constrained samples. These uses extend molecular analysis to specimens that may not provide enough material for direct examination.
Specificity and quantitative accuracy depend on controlling contamination and reaction conditions. Unwanted contaminating nucleic acids can generate products that are mistaken for the target, while poorly controlled conditions can distort how efficiently the sequence is copied or detected. Careful controls therefore matter most when comparing RNA amounts or interpreting weak signals from limited biological samples.