Rna Purification
Rna 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.
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
06:21
Depletion of Ribosomal RNA for Mosquito Gut Metagenomic RNA-seq
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Cited by 20 •
2013
A ribosomal RNA (rRNA) depletion protocol was developed to enrich messenger RNA (mRNA) for RNA-seq of the mosquito gut metatranscriptome. Sample specific rRNA probes, which were used to remove rRNA via subtraction, were created from the mosquito and its gut microbes. Performance of the protocol can result in the removal of approximately 90-99% of rRNA.
FAQs
RNA purification requires protection from ribonucleases because these enzymes can compromise the RNA being isolated. Lysis is therefore performed under conditions designed to inhibit their activity, helping preserve RNA molecules during subsequent separation and capture. Maintaining this protection supports recovery of RNA that is suitable for accurate downstream gene-expression and transcriptome analyses.
Removing proteins, DNA, lipids, and other cellular contaminants is essential because these substances can interfere with molecular analysis. A preparation may contain RNA yet still perform poorly if residual material disrupts downstream reactions or affects measurement. The purification process therefore emphasizes cleaner RNA recovery, improving the reliability and reproducibility of experiments that examine gene expression or RNA populations.
Chemical precipitation, organic extraction, and silica-based membrane capture all provide routes for selectively recovering RNA after cell lysis. These approaches differ in how RNA is separated from cellular material and contaminants. The choice of approach changes the capture stage of purification, while washing and elution produce the preparation used for later analysis.
RNA quality and integrity influence how accurately and reproducibly downstream experiments reflect the original biological sample. Damaged or otherwise compromised RNA can undermine gene-expression measurements, reverse transcription, quantitative PCR, RNA sequencing, or transcriptome analysis. For this reason, purification is not judged only by obtaining RNA, but also by preserving a preparation suitable for the intended assay.
A typical workflow begins with cell lysis under ribonuclease-inhibiting conditions. Cellular debris is then separated, after which RNA is selectively captured by chemical precipitation, organic extraction, or a silica-based membrane. Washing removes remaining unwanted material, and elution yields the RNA preparation used in downstream biology experiments.
Purified RNA provides the starting material for reverse transcription, quantitative PCR, RNA sequencing, and transcriptome analysis. These applications use the recovered RNA to investigate gene expression or broader RNA populations. Because the quality and integrity of the preparation affect accuracy and reproducibility, purification functions as a critical upstream step linking biological samples to dependable molecular measurements.