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Updated: Jun 3, 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

Isolation of plant nuclei.

R D Henfrey¹, R J Slater

¹Division of Biology and Environmental Sciences, The Hatfield Polytechnic, Hertfordshire, UK.

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

|March 23, 2011

Summary

This study details a method for isolating intact plant nuclei, crucial for gene expression research. Percoll gradient purification enhances nuclear preparations by removing contaminants and reducing ribonuclease activity, optimizing them for transcription studies.

Area of Science:

Plant molecular biology
Gene expression mechanisms
Nuclear biochemistry

Background:

Isolated plant nuclei are essential for studying gene expression control.
Applications include analyzing nuclear components, RNA processing, and transcription rates.
Existing methods require refinement for optimal experimental outcomes.

Purpose of the Study:

To describe a method for isolating crude plant nuclei.
To present a protocol for purifying nuclei using Percoll gradients.
To optimize nuclear preparations for gene expression and transcription studies.

Main Methods:

Isolation of crude plant nuclei from plant tissues.
Purification of nuclei using a discontinuous Percoll gradient.

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Assessment of nuclear integrity and purity post-isolation.

Main Results:

A reliable method for obtaining intact crude plant nuclei was established.
Percoll gradient centrifugation effectively removed cytoplasmic contaminants like starch grains.
The Percoll step was shown to reduce associated ribonuclease activity in nuclear preparations.

Conclusions:

The described method provides a robust approach for plant nuclei isolation.
Percoll purification is recommended for transcription experiments due to reduced contaminants and RNase activity.
Crude nuclei preparations may suffice for specific applications like initial RNA polymerase assays or component isolation.