Column-based Rna Extraction
Column-based Rna Extraction - Related Videos
02:59
Extraction of Viral RNA from Surface-Bound Virions Using a Silica Spin Column
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Source: Klaus, J. P. et al. Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment. J. Vis. Exp. (2016)This video demonstrates the extraction of viral RNA from surface-bound virions on epithelial cells using silica spin columns. The method includes chemical lysis, RNA stabilization, and membrane-based purification for downstream quantification of virus-cell attachment.
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Medium-Throughput Column DNA Extraction Using 24-Well Racks
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This protocol describes a cost-effective method to accelerate plasmid DNA extraction. By utilizing custom 3D-printed racks compatible with standard swinging-bucket plate rotors, researchers can process up to 48 spin columns with caps or up to 116 spin columns without caps simultaneously, significantly reducing handling time compared to manual microcentrifuge methods.
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RNA Extraction Assay: A Method to Extract RNA from miRNA Transfected Lung Cancer Cells
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This video describes the RNA extraction protocol from miRNA transfected lung cancer cells. This approach can be used to identify a combinatorial miRNA treatment activity in halting the cell cycle and angiogenesis.
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RNA Extraction from Neuroprecursor Cells Using the Bio-Rad Total RNA Kit
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09:18
Purification of a Total Lipid Extract with Column Chromatography
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2023
Source: Laboratory of Jeff Salacup - University of Massachusetts Amherst The product of an organic solvent extraction, a total lipid extract (TLE), is often a complex mixture of hundreds, if not thousands, of different compounds. The researcher is often only interested in a handful of compounds. The compounds of interest may belong to one of several classes of compounds, such as alkanes, ketones, alcohols, or acids (Figure 1), and it may be useful to remove the compound classes to which it does...
FAQs
Chaotropic salts serve two linked purposes during column-based RNA extraction. They disrupt cellular structures, helping release RNA from biological material, and inactivate RNases, enzymes that can degrade RNA. This chemical environment is essential because it protects the target molecule while preparing the sample for subsequent binding to the silica-based membrane.
High-salt conditions promote RNA binding to the silica-based membrane, allowing the target molecule to remain associated while other sample components can be removed. During elution, a low-salt solution reverses the conditions that favor retention, releasing purified RNA from the membrane. This change in chemical environment connects selective capture with recovery for downstream analysis.
Wash steps remove contaminants that remain after RNA has bound to the membrane. Their role is distinct from the initial chemical disruption and the final elution: washing cleans the retained material before recovery. Effective contaminant removal helps produce purified RNA suitable for downstream procedures such as reverse-transcription PCR, quantitative gene-expression analysis, and RNA sequencing.
A typical workflow starts with a biological sample, applies chemical conditions that disrupt the sample and suppress RNase activity, and then places the prepared material under high-salt conditions for membrane binding. Wash steps remove remaining contaminants, after which low-salt elution recovers the RNA. This sequence provides a reproducible path from sample preparation to purified material.
The approach can be used with tissues, cultured cells, blood, and microbial samples. This range makes the column workflow relevant to investigations involving different biological systems rather than a single sample type. Regardless of source, the procedure is designed to isolate RNA that can support analyses of gene expression and other RNA-dependent processes.
Purified RNA can be used for reverse-transcription PCR, quantitative gene-expression analysis, and RNA sequencing. These applications allow researchers to examine RNA-dependent processes, cellular responses, development, and disease-related changes. In biology, the extracted material therefore connects sample-level isolation with measurements of how gene activity and cellular states vary across experimental conditions.