Rna Amplification Method
Rna Amplification Method - 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.
09:06
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
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Cited by 1 •
2025
The protocol describes a tandem immunofluorescence hybridization assay with formalin-fixed paraffin-embedded tissue to detect and quantify microRNA and multiplexed proteins simultaneously.
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.
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.
FAQs
Reverse transcriptase provides the bridge between the original RNA and the amplified material by converting RNA into complementary DNA, or cDNA. This step creates a DNA form that sequence-specific primers can direct through repeated copying. In practice, it allows RNA abundance to be examined through an amplification workflow rather than relying on the original transcript quantity alone.
Sequence-specific primers determine which RNA-derived sequences are copied during amplification. Their guidance helps focus repeated enzyme-driven copying on transcripts of interest instead of treating all genetic material identically. This selectivity is especially useful when researchers want to examine particular neuronal, glial, developmental, disease-related, or injury-associated transcripts in a complex brain-tissue sample.
RNA amplification increases the amount of RNA-derived material available for detection and analysis, so low-abundance transcripts become more measurable. This greater sensitivity is important when researchers have limited sample material or when RNA is degraded. In neuroscience, the approach can therefore extend transcript analysis to brain samples that may not provide abundant, easily measured RNA.
Both approaches use reverse transcription and primer-guided copying, but they differ in the reaction condition used to repeat amplification. Thermal-cycling formats drive successive copying through changes in temperature, whereas isothermal formats maintain amplification under a constant-temperature condition. The choice therefore describes how the reaction is run, while primer specificity determines the target.
Researchers begin with RNA from a brain or other neuroscience sample, use reverse transcriptase to generate cDNA, and then apply sequence-specific primers in a thermal-cycling or isothermal amplification reaction. The resulting amplified RNA-derived material can be analyzed to examine transcript abundance. This workflow is valuable when the starting sample contains little available RNA.
In neuroscience, amplified material can support gene-expression profiling across brain tissue and focused analysis of neuronal or glial transcripts. Researchers can examine molecular changes associated with development, disease, or injury, using the increased analytical sensitivity to study patterns that may be difficult to detect from the original sample alone.
The analysis can reveal changes in transcript levels and broader gene-expression patterns, providing molecular evidence for differences in brain states or biological conditions. Such results may help identify candidate biomarkers or pathways for further investigation. Amplification improves access to these signals, but the resulting candidates remain subjects for additional study rather than established conclusions.