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DNA Isolation

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...

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Updated: Apr 22, 2026

Nuclei Isolation from Whole Tissue using a Detergent and Enzyme-Free Method

Nuclei Isolation from Whole Tissue using a Detergent and Enzyme-Free Method

Published on: June 24, 2020

A new rapid method for isolating nucleoli.

Zhou Fang Li¹, Yun Wah Lam

¹Department of Biology, South University of Science and Technology of China, 1088 Xueyuan Blvd., Nanshan, Shenzhen, Guangdong, P.R. China, zhoufang_li@hotmail.com.

Methods in Molecular Biology (Clifton, N.J.)

|October 15, 2014

Summary

This summary is machine-generated.

A new, rapid nucleolar isolation method uses snap-freezing and sonication for cultured cells. This technique enhances proteomic studies of the nucleolus (the cell

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Last Updated: Apr 22, 2026

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Area of Science:

Cell Biology
Proteomics
Subcellular Organelles

Background:

The nucleolus is a key cellular organelle, extensively studied using proteomic and live-cell imaging techniques.
Current methods for isolating nucleoli are often inefficient, requiring large cell numbers and complex procedures.
These limitations impede the detailed analysis of nucleolar dynamics and protein composition.

Purpose of the Study:

To develop and present an improved, rapid method for isolating nucleoli from cultured adherent cells.
To overcome the limitations of existing nucleolar isolation protocols.
To facilitate high-resolution proteomic studies of dynamic events within the nucleolus.

Main Methods:

A novel protocol involving snap-freezing of cultured adherent cells.
Direct sonication followed by centrifugation onto a sucrose cushion for nucleoli isolation.
The entire procedure is optimized for speed and efficiency, yielding results within 20 minutes.

Main Results:

Achieved rapid isolation of nucleoli in as little as 20 minutes.
The method provides a high yield, reducing the required starting material.
Preserves the integrity of nucleoli for accurate biochemical and proteomic analysis.
Enables the capture of transient biochemical changes by allowing precise time-point cell fixation.

Conclusions:

The developed protocol offers a significantly faster and more efficient approach to nucleolar isolation.
This method is ideal for proteomic studies investigating dynamic changes in nucleolar composition.
Enhances the understanding of mammalian cell biology through improved nucleolar research capabilities.