Acid Phenol Extraction
Acid Phenol Extraction - Related Videos
02:36
Acidity and Basicity of Alcohols and Phenols
0 Views •
2025
Like water, alcohols are weak acids and bases. This is attributed to the polarization of the O–H bond making the hydrogen partially positive. Moreover, the electron pairs on the oxygen atom of alcohol make it both basic and nucleophilic. Protonation of an alcohol converts hydroxide, a poor leaving group, into water—a good one. The two acid–base equilibria corresponding to ethanol are depicted below. Figure 1. Loss of proton Figure 2. Gain of proton Methanol (pKa = 15.5) is the only alcohol...
05:31
Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
0 Views •
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.
12:06
Synthesis of High Purity Nonsymmetric Dialkylphosphinic Acid Extractants
0 Views •
Cited by 6 •
2017
A protocol for the synthesis of high purity nonsymmetric dialkylphosphinic acid extractants is presented, taking (2,3-dimethylbutyl)(2,4,4'-trimethylpentyl)phosphinic acid as an example.
12:37
Efficient Nucleic Acid Extraction and 16S rRNA Gene Sequencing for Bacterial Community Characterization
0 Views •
Cited by 42 •
2016
We describe an efficient, robust, and cost effective method for extracting nucleic acid from swabs for characterization of bacterial communities using 16S rRNA gene amplicon sequencing. The method allows for a common processing approach for multiple sample types and accommodates a number of downstream analytic processes.
05:45
Extraction of Short-chain Fatty Acids from Mouse Feces Using A Two-phase Acid–Solvent Method
0 Views •
2026
This protocol describes a two-phase acid–solvent extraction method for isolating short-chain fatty acids from low-input mouse fecal samples without derivatization.
FAQs
The acidic solvent system changes the partitioning behavior of nucleic acids during phase separation. Under low-pH conditions, RNA remains primarily in the aqueous phase, while DNA is directed toward the interphase or organic phase. This distinction allows researchers to enrich RNA selectively, making the method useful when downstream analysis requires RNA rather than a mixture of nucleic acids.
Phenol denatures proteins, disrupting their native structure and promoting their movement away from the aqueous nucleic-acid fraction. After centrifugation, denatured proteins accumulate toward the interphase or organic phase, while RNA remains mainly aqueous under acidic conditions. This chemical and physical separation reduces protein contamination before the RNA-containing phase is collected.
The interphase and organic phase contain material that should generally remain separate from the RNA-containing aqueous fraction. DNA and denatured proteins partition toward these regions under the acidic conditions used. Disturbing the boundary can transfer unwanted components into the aqueous sample, reducing purity and potentially affecting later gene-expression analysis or reverse transcription.
The acidic condition, the effectiveness of protein denaturation, and the clarity of phase separation all influence the resulting RNA preparation. Inadequate separation can carry DNA or proteins into the aqueous fraction, while residual phenol can compromise sample quality. Careful handling of the phase boundary and thorough removal of phenol are therefore central to obtaining functional RNA.
The workflow begins by exposing cellular or tissue material to an acidic phenol-based solvent system, allowing proteins and other components to separate from nucleic acids. Centrifugation then produces aqueous, interphase, and organic regions. The RNA-containing aqueous phase is recovered while avoiding the boundary, followed by removal of phenol to prepare the sample for downstream use.
This approach is useful when researchers need RNA isolated from cells or tissues for gene-expression analysis, reverse transcription, or related molecular biology applications. Its value comes from combining protein denaturation with acidic phase partitioning, which enriches RNA in the aqueous fraction. The resulting preparation can support biochemical studies that depend on clean, functional RNA.