Related Experiment Video
Updated: Jun 3, 2026
07:00
Nuclei Isolation from Whole Tissue using a Detergent and Enzyme-Free Method
Published on: June 24, 2020
Isolation of plant nuclei
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.
- 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.
Related Concept Videos
01:24DNA 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...
01:34DNA Isolation
DNA from cells is required for many biotechnology and research applications, such as molecular cloning. To remove and purify DNA from cells, researchers use various methods of DNA extraction. While the specifics of different protocols may vary, some general concepts underlie the process of DNA extraction.
01:20Overview 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.
01:24Techniques 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...