Mrna Duplex Dissociation

mRNA duplex dissociation is the separation of two complementary messenger RNA strands or an mRNA strand and its complementary partner, a process that reveals how RNA stability influences gene regulation and molecular recognition. Dissociation occurs when heat, changes in ionic strength, or chemical denaturants weaken hydrogen bonding and base-stacking interactions, shifting the duplex toward single-stranded RNA; the transition depends on sequence, length, mismatches, and surrounding conditions. In biochemistry, measuring dissociation behavior helps characterize RNA hybridization, melting temperature, secondary structure, and binding kinetics. These insights support RNA assay design, probe development, therapeutics, and studies of post-transcriptional regulation.

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JoVE Core - Molecular Biology

Nuclear Export of mRNA

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2020

Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...

Research

JoVE Journal - Biology

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2012

Crystal structure of protein–DNA complexes can provide insight into protein function, mechanism, as well as, the nature of the specific interaction. Here, we report how to optimize the length, sequence and ends of duplex DNA for co-crystallization with Escherichia coli SeqA, a negative regulator of replication initiation.

Cell-Free In Vitro mRNA Translation In Drosophila Lysates

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2026

Here, we provide details on how to prepare lysates and perform in vitro translation using Drosophila embryos.

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2025

In this video, we demonstrate the procedure to determine the helicase activity of a target protein to unwind the biotin-labeled dsRNA substrate. The activity of the enzyme was identified by analyzing the electrophoretic mobility shift, followed by a chemiluminescence assay using chemiluminescent enzyme-conjugated streptavidin.

Duplex Digital PCR for Simultaneous Quantification of Dual Genetic Markers

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2025

In this video, we demonstrate the duplex digital PCR (ddPCR) technique — a modification of the traditional PCR technique that is useful in detecting two different genetic markers simultaneously. A single PCR reaction is partitioned into nanoliter-sized emulsified droplets that are independently amplified, and the detection of differently colored fluorescence amplification signals from the fraction of droplets is used to compute the initial concentration of the target sequences.

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