Abstract
This paper describes a modified FPG technique for differentiation of sister chromatids and for the study of sister chromatid exchanges in bean (Vicia faba) root tips. Roots were grown for one round of DNA replication in the presence of 5-bromodeoxyuridine and for a second round in the presence of thymidine. RNase-treated squash preparations of root tips grown in this way were stained with the fluorochrome compound “33258 Hoechst”, stored for 4 days at 4°C in 0.5×SSC, heated for 60 minutes at 55°C and stained for 6 to 7 minutes in a solution consisting of 3% Giemsa (Gurr's R66) in a 0.017 M phosphate buffer, pH 6.8. Chromatids with unsubstituted DNA strands (TT) stained darkly, whereas those containing 5-bromouracil in place of thymine in one of the strands of the DNA duplex (TB) stained weakly. Sister chromatid exchanges were observed and their frequency was proportional to chromosome length: 3.1 per M chromosome and 1.4 per S chromosome, giving a mean number of 20.6 exchanges per cell. If based on DNA content per cell, this frequency is in good agreement with those found by other authors in other materials. The technique should reveal single-strand exchanges between DNA double helices in sister chromatids if they occur; such exchanges were not observed, however.
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Kihlman, B.A., Kronborg, D. Sister chromatid exchanges in Vicia faba . Chromosoma 51, 1–10 (1975). https://doi.org/10.1007/BF00285801
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DOI: https://doi.org/10.1007/BF00285801