Rnase H
Rnase H - Related Videos
10:51
Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo
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Cited by 17 •
2014
We here describe a fluorescence based primer extension method to determine transcriptional starting points from bacterial transcripts and RNA processing in vivo using an automated gel sequencer.
08:49
Exosomal miRNA Analysis in Non-small Cell Lung Cancer (NSCLC) Patients' Plasma Through qPCR: A Feasible Liquid Biopsy Tool
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Cited by 59 •
2016
This protocol describes the feasibility to perform miRNA profiling in exosomes, released in plasma of NSCLC patients, through a commercial exosome isolation kit with Proteinase K and RNAse treatments, in order to avoid circulating miRNAs contamination and evaluate their biomarker features in NSCLC.
09:29
Stereotactic Atlas-Guided Laser Capture Microdissection of Brain Regions Affected by Traumatic Injury
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Cited by 3 •
2017
We describe the use of laser capture microdissection to obtain samples of distinct cell populations from different brain regions for gene and microRNA analysis. This technique allows the study of differential effects of traumatic brain injury in specific regions of the rat brain.
14:32
Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates
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Cited by 1 •
2016
This protocol describes the required steps to execute in vitro and in vivo deacetylation assays in order to establish the role of proteins as specific deacetylation substrates for sirtuins and further study the role of reversible - lysine acetylation as a post-translational modification.
02:42
Immunohistochemistry of Mouse Brain Tissue Sections for Targeting Dopamine Neurons
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2025
Source: Kummari, E., et. al., Laser Capture Microdissection - A Demonstration of the Isolation of Individual Dopamine Neurons and the Entire Ventral Tegmental Area., J. Vis. Exp. (2015)In this video, we demonstrate the immunohistochemistry protocol for dopamine neurons in brain tissue sections for subsequent laser capture microdissection.
FAQs
RNase H activity depends on divalent metal ions, especially magnesium or manganese. These ions activate water molecules for hydrolysis, the chemical cleavage of RNA phosphodiester bonds. Because the metal-dependent reaction targets the RNA strand within a hybrid, it supports selective removal while preserving the DNA strand for subsequent molecular processes.
The decisive substrate feature is an RNA-DNA hybrid, not simply the presence of RNA or DNA. RNase H preferentially hydrolyzes RNA phosphodiester bonds in this paired structure, whereas the DNA strand remains largely intact. That distinction lets investigators remove or process the RNA component without broadly destroying the associated DNA.
Selective cleavage prevents RNA-DNA hybrids from being handled as though both strands were equally degradable. By removing RNA while retaining DNA, RNase H contributes to controlled nucleic acid metabolism and genome integrity. This activity is particularly relevant when hybrids arise during transcription or reverse transcription, where unmanaged structures could complicate normal genetic processes.
During DNA replication, RNase H removes RNA primers after they have helped initiate DNA synthesis. Clearing these RNA segments allows the newly made DNA to remain as the relevant nucleic acid product rather than as a mixed primer-containing structure. This function connects hybrid-specific RNA degradation directly to completion and maintenance of replicated genetic material.
During transcription and reverse transcription, RNA-DNA hybrids can form as part of nucleic acid processing. RNase H activity helps manage these structures, supporting genome integrity and broader nucleic acid metabolism. In reverse transcription, the same activity is relevant to retroviral replication, linking basic hybrid processing with studies of retroviral biology.
Controlled hybrid degradation is useful when an experiment must remove an RNA component while retaining associated DNA. RNase H supports molecular biology methods that depend on selective hybrid processing and provides a basis for antisense research. Its metal-ion requirement and RNA-DNA substrate preference are central variables when designing or interpreting these applications.