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Continuous Percoll Gradient Using Swing Out Bucket Rotor

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Isolation of Highly Purified, Intact, and Functional Mitochondria from Potato Tubers Using a Two-in-One Percoll Density Gradient

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Part of the Methods in Molecular Biology book series (MIMB,volume 2363)

Abstract

The isolation of mitochondria from potato tubers (Solanum tuberosum L.) is described, but the methodology can easily be adapted to other storage tissues. After homogenization of the tissue, filtration and differential centrifugation, the key step is a Percoll density gradient centrifugation. The Percoll gradient contains two parts: a bottom part containing Percoll in 0.3 M sucrose, and a slightly less dense top part containing Percoll in 0.3 M mannitol. After centrifugation, a density gradient is formed that is almost linear in the central part, and this is where the band containing the purified intact mitochondria is formed. This method makes it possible to process large amounts of plant material (2–6 kg) and saves at least 1.5 h on the preparation time compared to methods where two consecutive purification methods are used. Nonetheless, it yields large amounts of mitochondria (50–125 mg protein) of very high purity, intactness and functionality.

Key words

  • Amyloplast envelope
  • Mitochondria
  • Percoll gradient purification
  • Peroxisomes
  • Potato tubers
  • Solanum tuberosum
  • Starch
Fig. 1

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Author information

Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Aarhus University, Slagelse, Denmark

    Ian Max Møller

  2. Department of Biology, Lund University, Lund, Sweden

    Allan G. Rasmusson

Corresponding author

Correspondence to Ian Max Møller .

Editor information

Editors and Affiliations

  1. Department of Biology, Lund University, Lund, Sweden

    Dr. Olivier Van Aken

  2. Department of Biology, Lund University, Lund, Sweden

    Prof. Allan G. Rasmusson

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Møller, I.M., Rasmusson, A.G. (2022). Isolation of Highly Purified, Intact, and Functional Mitochondria from Potato Tubers Using a Two-in-One Percoll Density Gradient. In: Van Aken, O., Rasmusson, A.G. (eds) Plant Mitochondria. Methods in Molecular Biology, vol 2363. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1653-6_4

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  • DOI : https://doi.org/10.1007/978-1-0716-1653-6_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1652-9

  • Online ISBN: 978-1-0716-1653-6

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