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Technological exploration of BAC-FISH on mitotic
chromosomes of maize |
| TAO Yongsheng1, ZHANG Zuxin2, CHEN Yonglin3, LI Lijia3, ZHENG Yonglian4 |
| 1.National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University；College of Agronomy, Hebei Agricultural University; 2.College of Agronomy, Hebei Agricultural University; 3.Key Laboratory of MOE for Plant Developmental Biology, Wuhan University; 4.National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University |
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Abstract The rice BAC-DNA was used as probes and fluorescence in situ hybridization (FISH) was applied to the interphase and metaphase mitotic chromosomes of maize. To optimize the BAC-FISH technique, we respectively assayed the effect of several factors, including maize or rice genomic Cot DNA used as blocking reagent of DNA, washing temperatures and FAD concentration in the washing buffer and in the hybrid solution. The results show that Cot DNA of maize genome blocked the repetitive sequence of the rice BAC-DNA when the Cot value was below 50. Meanwhile, it was necessary to adjust the Cot value according to the different probes and their ratios. Decreasing the concentration of FAD in the hybridization mixtures, adjusting the washing rate after hybridization, and most especially, blocking the rice-specific repetitive sequences of BAC-DNA could improve the positive signals of BAC-FISH.
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Issue Date: 05 December 2008
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