Alkaline Lysis Method
Alkaline Lysis Method - Related Videos
07:46
On-site DNA Detection of Trypanosomatid Parasites and Nosema ceranae Through Alkaline Lysis Coupled to RPA/CRISPR/Cas12a System
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2025
Here, we describe a simple and quick procedure for detecting DNA from bee pathogens, such as Lotmaria passim and Nosema ceranae, using an amplification-ready cell lysis, recombinase polymerase amplification, and CRISPR/Cas12a assays.
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.
09:49
A Simple Red Blood Cell Lysis Method for the Establishment of B Lymphoblastoid Cell Lines
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Cited by 1 •
2017
A simple red blood cell lysis method for establishing B-LCLs was developed with high immortalization efficiency, requiring a small amount of blood and saving time from initiation to cryopreservation.
03:12
A Procedure for Bacterial Harvesting and Mechanical Lysis
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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 a step-by-step procedure for harvesting bacterial cells and mechanically lysing them to release intracellular contents.
01:32
Tissue Homogenization and Cell Lysis
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2023
Tissue homogenization involves disintegrating tissue architecture and lysing cells, and is an early step in isolating and analyzing cellular components. The method used for homogenization depends on the sample type, the amount of sample available, the analyte to be obtained, and the sensitivity of the method. These methods are broadly classified as mechanical and non-mechanical methods. Mechanical methods of tissue homogenization These methods rely on applying external physical force to disrupt...
FAQs
Sodium hydroxide creates the alkaline conditions that denature proteins and nucleic acids, while SDS is a detergent that disrupts bacterial cell membranes. Using both reagents first breaks open the cells and exposes their contents for selective separation during neutralization. This coordinated chemical treatment is central to releasing plasmid DNA from the bacterial cell material.
Potassium acetate neutralizes the alkaline mixture after cell disruption. Under these conditions, plasmid DNA can renature, whereas genomic DNA, denatured proteins, and cell debris precipitate. This difference in behavior produces a plasmid-containing liquid phase and a precipitated fraction, allowing the desired DNA to remain in solution for subsequent recovery.
Resuspension places the bacterial cells into a prepared, evenly distributed starting suspension before sodium hydroxide and SDS are added. This establishes the initial condition for membrane disruption and molecular denaturation. In the overall workflow, the step connects intact-cell handling with chemical lysis and helps ensure that the cellular material enters the separation process together.
Centrifugation separates the precipitated genomic DNA, proteins, and cell debris from the liquid that contains plasmid DNA. The resulting physical separation produces a clearer plasmid-containing solution rather than leaving the target material mixed with the insoluble fraction. That solution can then undergo further purification before being used in downstream molecular biology work.
Researchers choose this method when they need plasmid DNA from bacterial cells for genetic research. The isolated material can support cloning, recombinant DNA workflows, bacterial transformation, sequencing, and analysis of engineered genetic constructs. Its value lies in connecting bacterial culture material with downstream experiments that require access to plasmid-borne genetic information.
After separation and any further purification, the plasmid-containing solution provides DNA for examining engineered genetic constructs. Researchers can use it in cloning and recombinant DNA workflows, introduce it during bacterial transformation, or analyze its sequence. These applications make the method relevant for following how designed genetic material is assembled, propagated, and evaluated.