Rna Sample Purification
Rna Sample Purification - Related Videos
11:09
Chromatin Isolation by RNA Purification (ChIRP)
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Cited by 380 •
2012
ChIRP is a novel and rapid technique to map genomic binding sites of long noncoding RNAs (lncRNAs). The method takes advantage of the specificity of anti-sense tiling oligonucleotides to allow the enumeration of lncRNA-bound genomic sites.
02:30
Isolation of Small Regulatory RNA–Bound Bacterial Target RNA Using Affinity Purification
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2025
Source: Mercier, N., et. al., MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria. J. Vis. Exp. (2021)This video demonstrates the isolation of MS2-tagged small regulatory RNA (sRNA) bound to its bacterial target RNA using affinity purification with a maltose-binding protein (MBP)–MS2 coat protein fusion immobilized on amylose resin.
10:42
RNA Purification from Intracellularly Grown Listeria monocytogenes in Macrophage Cells
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Cited by 11 •
2016
Here we describe a method for bacterial RNA isolation from Listeria monocytogenes bacteria growing inside murine macrophages. This technique can be used with other intracellular pathogens and mammalian host cells.
12:04
RNA Analysis of Environmental Samples Using RT-PCR
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2023
Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - The University of Arizona Demonstrating Author: Bradley Schmitz Reverse transcription-polymerase chain reaction (RT-PCR) involves the same process as conventional PCR — cycling temperature to amplify nucleic acids. However, while conventional PCR only amplifies deoxyribonucleic acids (DNA), RT-PCR enables the amplification of ribonucleic acids (RNA) through the formation of complementary DNA (cDNA). This enables RNA-based organisms...
11:00
Genotyping of Plant and Animal Samples without Prior DNA Purification
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Cited by 14 •
2012
The Direct PCR approach presented here facilitates PCR amplification directly from small amounts of unpurified plant and animal tissue.
FAQs
The matrix provides the selective separation step: RNA binds to it under the purification conditions, while subsequent washing removes remaining proteins, DNA, lipids, and other contaminants. Elution then releases the retained RNA into a usable fraction. This sequence connects chemical separation with recovery of material suitable for expression measurements, reverse transcription, transcript profiling, or sequencing.
RNase inhibition helps preserve RNA integrity during processing. Cell or tissue lysis exposes cellular contents, so the workflow pairs disruption with conditions that limit RNase activity before RNA is bound, washed, and eluted. Maintaining intact RNA improves the reliability of downstream transcript profiling, gene-expression comparisons, and investigations of molecular differences between cellular states.
Proteins, DNA, lipids, and other contaminants are removed so the recovered fraction is enriched for RNA rather than an unseparated cellular mixture. This cleanup supports downstream reverse transcription, transcript profiling, sequencing, and gene-expression measurements. In genetics, obtaining a cleaner RNA preparation helps researchers examine regulation and molecular changes with greater confidence in the sample material.
The workflow begins with cell or tissue lysis, accompanied by measures that inhibit RNases. RNA is then selectively bound to a purification matrix, followed by washing to remove unwanted material. Finally, controlled elution recovers the RNA. Preserving RNA integrity across these stages is essential before applying the preparation to genetic analyses.
Lysis releases the contents of cells or tissues so RNA becomes accessible for purification. Binding transfers RNA to the purification matrix, while washing removes proteins, DNA, lipids, and other residual material. Elution recovers the retained RNA after cleanup. Treating these stages as separate functions helps maintain a preparation appropriate for downstream analysis.
Purified RNA provides material for measuring gene expression, profiling transcripts, performing reverse transcription, and conducting sequencing. These applications allow researchers to compare cellular states, investigate gene regulation, and identify molecular changes. The purification step is therefore important because the quality and cleanliness of the recovered RNA directly support interpretation of downstream genetic measurements.