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Related Concept Videos

DNA Isolation01:34

DNA 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.
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...
DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
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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.
Subcellular Fractionation01:32

Subcellular Fractionation

The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
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Related Experiment Video

Updated: Jul 11, 2026

On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids
10:32

On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids

Published on: March 2, 2012

RNA isolation and fractionation with compaction agents.

J C Murphy1, G E Fox, R C Willson

  • 1Department of Chemical Engineering, University of Houston, 4800 Calhoun Avenue, Houston, TX 77204-4792, USA.

Analytical Biochemistry
|August 8, 2001
PubMed
Summary

Researchers developed a novel RNA isolation method using selective precipitation with compaction agents like hexammine cobalt. This technique efficiently separates total RNA, ribosomal RNA (rRNA), and low molecular weight RNA from bacterial lysates.

Keywords:
NASA Discipline Environmental HealthNon-NASA Center

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Purification

Background:

  • Traditional RNA isolation methods can be complex and time-consuming.
  • Bacterial lysates contain a mixture of RNA species requiring differential purification.
  • Compaction agents offer potential for selective nucleic acid precipitation.

Purpose of the Study:

  • To introduce and validate a new RNA isolation technique using selective precipitation.
  • To demonstrate the efficacy of hexammine cobalt and spermidine as compaction agents.
  • To enable fractionation of different RNA types from bacterial samples.

Main Methods:

  • Selective precipitation of total RNA using 3.5 mM hexammine cobalt.
  • Fractionation of ribosomal RNA (rRNA) from low molecular weight RNA using 2 mM hexammine cobalt.
  • Nondenaturing anion-exchange chromatography for resolving 5S, 16S, and 23S rRNA.
  • Second-stage precipitation at 8 mM hexammine cobalt for isolating low molecular weight RNA.
  • Application of the method to purified artificial stable RNA and expressed ribozymes.

Main Results:

  • Total RNA was selectively precipitated from bacterial lysates.
  • Ribosomal RNA (rRNA) was effectively fractionated from low molecular weight RNA.
  • Pure 5S rRNA and mixed 16S/23S rRNA were resolved using anion-exchange chromatography.
  • Low molecular weight RNA fraction was successfully isolated via precipitation.
  • The method proved effective for purifying artificial stable RNA and ribozymes.

Conclusions:

  • Selective precipitation by compaction agents provides an efficient approach for RNA isolation.
  • This method allows for the fractionation of diverse RNA species from bacterial sources.
  • The technique is versatile, applicable to both native and engineered RNA molecules.