Alkaline Extraction
Alkaline Extraction - Related Videos
10:38
In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage in Rat Liver
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Cited by 17 •
2016
The alkaline comet assay measures DNA strand breaks in eukaryotic cells. By adding an Endonuclease III or human 8-oxoguanine-DNA-N-glycosylase digestion step, the assay can efficiently detect oxidative DNA damage. We describe methods for using these assays to detect DNA damage in rat liver.
03:11
Quantification of Bioactive Type-I Interferons using a Secreted Embryonic Alkaline Phosphatase Reporter Assay
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2025
This video demonstrates secreted embryonic alkaline phosphatase assay using HEK-Blue interferon-α/β reporter cells to quantify the type-I interferons (IFNs) released following stimulation of plasmacytoid dendritic cells by HIV-1 Infected CD4+ T cells.
06:18
Assessing Lysosomal Alkalinization in the Intestine of Live Caenorhabditis elegans
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Cited by 6 •
2018
A step-by-step guide to probe loss of lysosomal acidity in the intestine of C. elegans using the pH-sensitive vital dye 5(6)-carboxy-2',7'-dichlorofluorescein diacetate...
02:55
Extraction - Concepts
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2020
Extraction Extraction is a common technique used in organic chemistry to isolate a target compound. In the extraction process, a solute is transferred from one phase to another to separate it from unreacted starting materials or impurities. Extraction is also used to facilitate the isolation of a solute from a reaction solvent that is difficult to remove by evaporation, such as a solvent with a high boiling point. Generally, there are three types of extractions. First, in solid-liquid...
06:42
Extraction of Biomarkers from Sediments - Accelerated Solvent Extraction
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2023
Source: Laboratory of Jeff Salacup - University of Massachusetts Amherst The distribution of a group of organic biomarkers called glycerol-dialkyl glycerol-tetraethers (GDGTs), produced by a suite of archaea and bacteria, were found in modern sediments to change in a predictable manner in response to air or water temperature1,2. Therefore, the distribution of these biomarkers in a sequence of sediments of known age can be used to reconstruct the evolution of air and/or water temperature on...
FAQs
Sodium hydroxide raises pH, changing the chemical behavior of cellular components, while detergent helps lyse cells and make intracellular contents accessible. Under these conditions, chromosomal DNA and proteins become denatured, meaning their molecular structures are disrupted. This combination creates the conditions needed to distinguish plasmid DNA from other cellular material during subsequent neutralization.
Neutralization reverses the extreme-pH conditions enough to promote renaturation of plasmid DNA, allowing complementary strands to reassociate. At the same time, cellular debris precipitates. This contrast supports separation of plasmid DNA from disrupted cellular material, because the DNA undergoes a different chemical response from the denatured components that form the precipitated debris.
High pH changes the ionization state of biomolecules, which alters their chemical interactions. It also disrupts noncovalent interactions, the weak attractions that help maintain associations among molecules and cellular structures. These changes explain why alkaline conditions can release target molecules and alter the behavior of DNA, proteins, and other sample components during extraction.
A plasmid preparation generally begins by exposing cells to an alkaline solution containing sodium hydroxide and detergent. The treatment lyses cells and denatures chromosomal DNA and proteins. A neutralization step follows, promoting plasmid DNA renaturation while cellular debris precipitates. This sequence uses controlled chemical changes to separate plasmid DNA from disrupted cellular material.
The method is useful when researchers need to release and separate target molecules from cells or complex sample materials through pH-dependent chemical behavior. In plasmid DNA preparation, the resulting isolation supports cloning, sequencing, and other molecular biology applications. Its value lies in combining relatively simple chemical treatment with separation of cellular components.
Its main product is isolated plasmid DNA rather than a complete mixture of cellular contents. Because the process separates plasmid DNA from denatured chromosomal DNA, proteins, and precipitated debris, the preparation can support downstream cloning, sequencing, and related molecular biology workflows. The outcome demonstrates how pH-dependent biomolecular behavior enables practical purification.