Nuclei Lysis
Nuclei Lysis - Related Videos
03:55
Isolation of Nuclei from Fresh Frozen Glioma Tissues: A Method to Obtain Intact Nuclei from Glioma Tumor Samples
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2023
This video describes the protocol to isolate single-nuclei from fresh frozen glioma tissue. The isolated single-nuclei can be used as specimens for sequencing techniques such as single-nuclei RNA (snRNA) sequencing and single-nuclei Assay for Transposase-Accessible Chromatin (snATAC) sequencing to obtain deeper insights into inter- and intra-tumor heterogeneity observed in glioma of the central nervous tissues.
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.
10:12
Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue
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Cited by 61 •
2012
Cardiac nuclei are isolated via density sedimentation and immunolabeled with antibodies against pericentriolar material 1 (PCM-1) to identify and sort cardiomyocyte nuclei by flow cytometry.
10:58
Neuronal Nuclei Isolation from Human Postmortem Brain Tissue
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Cited by 136 •
2008
The cellular heterogeneity of brain tissue poses a significant limitation for the study of epigenetic markings in chromatin because most assays lack single cell resolution. Neurons typically are intermingled with glia and other non-neuronal cells. We provide a protocol to extract and collect neuronal nuclei from human brain.
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
Hypotonic buffers create conditions that weaken the nuclear envelope, allowing water movement to contribute to nuclear swelling and rupture. This approach can release genomic DNA, chromatin, RNA, and nuclear proteins without relying solely on harsh mechanical force. Buffer conditions must still be controlled because excessive disruption may increase degradation or unwanted contamination in downstream biological analyses.
These approaches disrupt nuclei through different mechanisms. Detergents weaken membrane structures, enzymatic treatment helps break down structural components, and mechanical shearing applies physical force to rupture or fragment nuclear material. Researchers can use them individually or in combination, selecting conditions according to whether DNA, chromatin, RNA, or nuclear proteins must remain suitable for subsequent analysis.
Efficient rupture improves the release and recovery of nuclear molecules, but excessive disruption can promote degradation or carry unwanted cellular material into the preparation. Controlled conditions therefore support both sample quality and reproducibility. This balance is especially important when the recovered genomic DNA, chromatin, RNA, or nuclear proteins will undergo purification, molecular assays, or sequencing-related preparation.
The intended analyte determines which aspects of disruption require the most control. Genomic DNA and chromatin may require recovery of intact nuclear material, while RNA and nuclear proteins require conditions that limit degradation and preserve their analytical usefulness. Consequently, researchers adjust the lysis approach to the molecule being studied rather than treating every nuclear preparation identically.
A general workflow begins by exposing cells or nuclei to selected disruption conditions, such as a hypotonic buffer, detergent, enzymatic treatment, or mechanical shearing. The resulting preparation is then used for recovery or purification of the desired nuclear material. Throughout the process, researchers control disruption to improve yield, reduce contamination, and maintain consistency between samples.
Researchers use this step when experiments require access to nuclear contents rather than only intact cells. Supported applications include DNA and chromatin extraction, RNA or nuclear protein analysis, nucleic acid purification, and preparation for sequencing or other molecular assays. The method is therefore relevant across studies that examine genetic material, gene regulation, genome organization, or cellular function.
Nuclear preparations provide access to genomic DNA, chromatin, RNA, and nuclear proteins that underlie nuclear processes. Analyzing these components can support investigations of gene regulation, genome organization, and cellular function. Reliable lysis contributes to interpretable results by improving molecular recovery and reproducibility, while controlled conditions help limit degradation and unwanted contamination that could affect downstream analyses.