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Related Concept Videos

DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
Techniques for Isolation of Pure Cultures01:24

Techniques for Isolation of Pure Cultures

Microorganisms are routinely cultured in the laboratory using various techniques to isolate, grow, and quantify them for further study. These methods rely on inoculating microorganisms into a suitable growth medium under aseptic conditions to prevent contamination. Depending on the objective, inoculation can involve direct transfer or the use of diluted bacterial suspensions as the inoculum.Streak-Plate Method for IsolationThe streak-plate method is a common technique for obtaining pure...
Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.

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Related Experiment Video

Updated: Jun 19, 2026

Single-Copy Gene Locus Chromatin Purification in Saccharomyces cerevisiae
10:33

Single-Copy Gene Locus Chromatin Purification in Saccharomyces cerevisiae

Published on: November 17, 2023

Isolation of nuclei from yeast.

M M Bhargava1, H O Halvorson

  • 1Laboratory of Molecular Biology and the Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706.

The Journal of Cell Biology
|October 30, 2009
PubMed
Summary

A new method efficiently isolates yeast nuclei, yielding a high DNA/protein ratio. These isolated nuclei exhibit key structural features and maintain RNA synthesis capabilities, proving useful for molecular studies.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Efficient isolation of Saccharomyces cerevisiae nuclei is crucial for studying nuclear components and functions.
  • Previous methods may yield low quantities or compromised nuclear integrity.

Purpose of the Study:

  • To develop and describe a high-yield method for isolating nuclei from Saccharomyces cerevisiae.
  • To characterize the isolated nuclei and assess their suitability for further molecular analysis.

Main Methods:

  • A novel isolation procedure for Saccharomyces cerevisiae nuclei.
  • Microscopic examination (phase and electron microscopy) of isolated nuclei.
  • Investigation of optimal conditions for histone extraction and resolution.
  • Assay of RNA polymerase activity and in vitro RNA synthesis.

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Use of Time-Lapse Microscopy and Stage-Specific Nuclear Depletion of Proteins to Study Meiosis in S. cerevisiae
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Use of Time-Lapse Microscopy and Stage-Specific Nuclear Depletion of Proteins to Study Meiosis in S. cerevisiae

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Main Results:

  • High yield of isolated nuclei achieved.
  • Isolated nuclei show a 10-fold higher DNA/protein ratio compared to whole cells.
  • Microscopic analysis reveals double membranes, microtubules, and a characteristic crescent structure.
  • Active RNA polymerase (E.C. 2.7.7.6) confirmed, enabling in vitro RNA synthesis.
  • Nuclei are readily stainable with Giemsa's, Feulgen's, and acridine orange.

Conclusions:

  • The described method provides a reliable way to obtain pure, intact yeast nuclei in high yield.
  • Isolated nuclei are suitable for biochemical studies, including histone analysis and RNA synthesis.
  • These nuclei serve as a valuable resource for investigating yeast molecular biology and genetics.