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Flow sorting of wheat chromosome arms from the ditelosomic line 7BL
Authors:Dong-Wei Guo  You-Zhi Ma  Lian-Cheng Li  Yao-Feng Chen
Affiliation:(1) Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Institute of Crop Sciences, Chinese Academy of Agriculture Sciences, 100081 Beijing, China;(2) Agronomy College of Northwestern Sci-Tech University of Agriculture and Forestry, Yangling, 712100 Shannxi, China
Abstract:Flow cytometric analysis confirmed that root tip cells can be synchronized with 1.25 mM hydroxyurea (DNA synthesis inhibitor) for 12 h and 1 μM trifluralin (metaphase blocking reagent) treatment for 5 h. Chromosome suspensions prepared from homogenized tissue were suitable for chromosome sorting. A flow karyotypic histogram showed that the genome of common wheat (Triticum aestivum L.) ‘Chinese Spring’ was divided into 4 chromosome peaks, but the 7BL ditelosomic line had an additional chromosome peak. PCR amplification of sorted chromosome arms indicated that the extra chromosome peak consisted of 7BL telosomics. Some technical details of sample preparation and parameter setting for flow cytometric analysis are described.
Keywords:chromosome sorting  flow cytometry  flow karyotype  telosomic isolation
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