麦类作物体细胞中GISH的影响因素分析

Shi yan sheng wu xue bao Pub Date : 2005-04-01
Hiao Hu Lin, Xing Feng Li, Li Ming Wang, Wen Hui lu, Hong Gang Wang
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引用次数: 0

摘要

分析了影响基因组原位杂交(GISH)技术在小麦科作物(包括八倍体小黑麦、八倍体小黑麦、八倍体小黑麦和小麦-麦中间外源二体附加系)体细胞中的应用的因素。结果表明,理想的GISH效果是建立在高质量染色体制剂的基础上的,该染色体制剂含有更多的细胞有丝分裂中期,染色体分散良好,无异物。关键因素是适宜的探针DNA与阻断DNA的比例和洗脱条件的控制。此外,还对染色体丢失、异源染色体无杂交信号、杂交信号强或弱、杂交信号差或过丰(高杂交背景)、杂交信号噪声、杂交斑点等异常现象进行了初步分析。对这些问题进一步提出了建议和解决办法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Analysis of factors influencing GISH used in Triticeae crops' body cells].

Factors influencing genomic in situ hybridization (GISH) used in the body cells of Triticeae crops including Octoploid Tritileymus, Octoploid Trititrigia, Octoploid Tritelytrigia and wheat-Elytrigia intermedium alien disomic addition line were analyzed. The results indicated that ideal GISH effects were based on the high quality chromosome preparations containing more cell mitotic metaphase, better dispersed chromosomes and no foreign matter. The critical factors were suitable ratio of probe DNA to blocker DNA, and the control of elution condition. In addition, some abnormal phenomena, such as chromosome loss, no hybridization signal in alien chromosome, strong or weak hybridization signal, poor or overabundant hybridization signal (high hybridization background), noisy hybridization signal and hybridization spots, were preliminarily analyzed. Proposals and solutions for these problems were further brought forth.

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