The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.

We have developed a protocol which facilitates the isolation of nuclei containing mitotic spindles from the plasmodium of Physarum polycephalum. The nuclei are isolated in a spindle stabilization buffer (SSB) consisting of -4 M glycerol, 5 mM EGTA, 10 mM MgCl2, 10 mM PIPES and 0.1% (v/v) Triton X-10...

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Main Authors: Roobol, A, Paul, E, Gull, K
Format: Journal article
Language:English
Published: 1984
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author Roobol, A
Paul, E
Gull, K
author_facet Roobol, A
Paul, E
Gull, K
author_sort Roobol, A
collection OXFORD
description We have developed a protocol which facilitates the isolation of nuclei containing mitotic spindles from the plasmodium of Physarum polycephalum. The nuclei are isolated in a spindle stabilization buffer (SSB) consisting of -4 M glycerol, 5 mM EGTA, 10 mM MgCl2, 10 mM PIPES and 0.1% (v/v) Triton X-100 at pH 6.5. Isolation at 20 degrees C, chelation of Ca2+ and the inclusion of the 4 M glycerol were found to be essential for stabilization of the spindle microtubules. The method allows the isolation of clean nuclei in a reasonable yield. Nuclei were isolated at each of the main stages of mitosis (prophase, metaphase and anaphase) and transmission electron microscopy of the isolated nuclei revealed well preserved arrangements of spindle microtubules which were characteristic of the particular stage of mitosis.
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spelling oxford-uuid:ba6a803a-e70f-42b2-baf2-72a5e0d3b87c2022-03-27T05:09:42ZThe isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:ba6a803a-e70f-42b2-baf2-72a5e0d3b87cEnglishSymplectic Elements at Oxford1984Roobol, APaul, EGull, KWe have developed a protocol which facilitates the isolation of nuclei containing mitotic spindles from the plasmodium of Physarum polycephalum. The nuclei are isolated in a spindle stabilization buffer (SSB) consisting of -4 M glycerol, 5 mM EGTA, 10 mM MgCl2, 10 mM PIPES and 0.1% (v/v) Triton X-100 at pH 6.5. Isolation at 20 degrees C, chelation of Ca2+ and the inclusion of the 4 M glycerol were found to be essential for stabilization of the spindle microtubules. The method allows the isolation of clean nuclei in a reasonable yield. Nuclei were isolated at each of the main stages of mitosis (prophase, metaphase and anaphase) and transmission electron microscopy of the isolated nuclei revealed well preserved arrangements of spindle microtubules which were characteristic of the particular stage of mitosis.
spellingShingle Roobol, A
Paul, E
Gull, K
The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.
title The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.
title_full The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.
title_fullStr The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.
title_full_unstemmed The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.
title_short The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.
title_sort isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould physarum polycephalum
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