A standardized method for the three-way differential staining of plant chromosomes and the scoring of SCEs per cell cycle

We have made use of 2 alternative methodologies to obtain 3-way differential staining (TWD) in third-mitosis (M3) chromosomes of Allium cepa, which involve different uptakes of bromodeoxyuridine (BrdU) through 3 cell divisions, in order to evaluate the sister-chromatid exchanges (SCE) frequency on a...

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Bibliographic Details
Published inMutation research Vol. 216; no. 4; p. 203
Main Authors Escalza, P, Piñero, J, Cortés, F
Format Journal Article
LanguageEnglish
Published Netherlands 01.08.1989
Subjects
Allium - genetics
Bromodeoxyuridine
Cell Cycle
Chromosomes
DNA - drug effects
DNA - genetics
Floxuridine - pharmacology
Genetic Techniques
Sister Chromatid Exchange
Staining and Labeling
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Summary:We have made use of 2 alternative methodologies to obtain 3-way differential staining (TWD) in third-mitosis (M3) chromosomes of Allium cepa, which involve different uptakes of bromodeoxyuridine (BrdU) through 3 cell divisions, in order to evaluate the sister-chromatid exchanges (SCE) frequency on a per-cell-cycle basis. The main innovation introduced by us to the original protocols previously reported has been the use of 5-fluorodeoxyuridine (FdU) to inhibit endogenous thymidine (dT) synthesis. By using different [BrdU]:[dT] ratios in the presence of FdU the relative incorporation of BrdU into replicating DNA can be controlled. The use of 2 different approaches to obtain 3-way differentiation of sister chromatids allowed us to evaluate the role of BrdU in the induction of SCEs in our system. Both procedures rendered nearly 100% of M3 chromosomes showing TWD. An additional advantage of our methodologies is their high degree of reproducibility.
ISSN:0027-5107