Hot Phenol-sds Extraction
Hot Phenol-sds Extraction - Related Videos
05:31
Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
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Cited by 168 •
2012
We describe a modified hot aqueous-phenol extraction method for purifying lipopolysaccharide (LPS) from Gram-negative bacteria. Once extracted, the LPS can be subsequently analyzed by SDS-PAGE and visualized by direct staining or Western immunoblot.
09:23
Optimized Hot Phenol–Based RNA Extraction from Mycobacteria: A Robust Approach for Reliable Gene Expression Analysis
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2026
This study optimizes a Hot Phenol–based RNA extraction method for Mycobacterium, yielding high-quality RNA suitable for transcriptomic analyses. When benchmarked against TRIzol and RNeasy under identical conditions, the method provided higher yield, comparable integrity, and improved cost-efficiency, offering a practical alternative for large-scale gene expression studies.
03:22
SDS-PAGE Based Extraction of Extracellular Vesicles Associated Proteins: A Procedure to Extract Proteins from EVs and Prepare Them for In-Gel Digestion
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2023
In this video, we demonstrate the SDS-PAGE in-gel digestion method for protein extraction from extracellular vesicles or EVs. Once isolated, the obtained protein fragments can be used for proteomic analysis.
07:29
Separating Protein with SDS-PAGE
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2023
Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis, or SDS-PAGE, is a widely-used technique for separating mixtures of proteins based on their size and nothing else. SDS, an anionic detergent, is used to produce an even charge across the length of proteins that have been linearized. By first loading them into a gel made of polyacrylamide and then applying an electric field to the gel, SDS-coated proteins are then separated. The electric field acts as the driving force, drawing the SDS...
01:27
SDS-PAGE
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2023
Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field. A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
FAQs
SDS and hot phenol contribute different chemical actions during extraction. SDS disrupts cell membranes and denatures proteins, releasing nucleic acids from cellular material. Heated phenol then further unfolds protein complexes, helping separate those nucleic acids from proteins during centrifugation. The combined treatment therefore links membrane lysis, protein disruption, and phase-based recovery in one workflow.
Phase separation depends on partitioning the sample into distinct layers after treatment and centrifugation. The nucleic-acid-containing aqueous phase is recovered, while the separation step helps exclude material associated with the other phase. Careful identification and transfer of the aqueous fraction are therefore central to preserving the desired DNA or RNA for subsequent precipitation and analysis.
Protein denaturation matters because the method must separate nucleic acids from protein material. SDS disrupts membranes and denatures proteins, while hot phenol further unfolds protein complexes. These complementary effects promote transfer of nucleic acids into the aqueous phase, making phase separation more effective for recovering DNA or RNA for downstream work.
A basic workflow starts with detergent-mediated lysis, followed by heated phenol treatment and centrifugation to separate phases. The aqueous fraction is then recovered, and the nucleic acids are precipitated, washed, and analyzed. This sequence moves from cell disruption to chemical separation and finally cleanup, producing DNA or RNA preparations suitable for electrophoresis, amplification, or gene-expression analysis.
Recovered preparations can be examined by electrophoresis, used in amplification, or applied to gene-expression analysis. These downstream options allow investigators to inspect or use the isolated nucleic acids according to the experimental objective. Because the extraction yields DNA or RNA preparations, it can serve as a common upstream step for different molecular analyses in infection and immunology studies.
In immunology and infection research, the method can support separate but related questions about pathogen and host biology. DNA preparations may be used to study microbial genomes, while RNA preparations can support analysis of pathogen-associated transcripts or host responses. This makes the extraction relevant when nucleic acid recovery is needed to connect infection-related material with molecular measurements.