杂交鲍染色体GISH的优化-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Optimization of Genomic in situ Hybridization (GISH) for Hybrid Abalone

作者:: 刘富江1; 2; 朱齐春1; 2; 张健鹏1; 2; 蔡明夷1; 2; （1.集美大学水产学院，福建厦门 361021；2.农业农村部东海海水健康养殖重点实验室，福建厦门 361021）

Author(s):: LIU Fujiang1; 2; ZHU Qichun1; 2; ZHANG Jianpeng1; 2; CAI Mingyi1; 2; (1.Fisheries College,Jimei University,Xiamen 361021,China;2.The Key Laboratory of Healthy Mariculture for the East China Sea,Ministry of Agriculture and Rural Affairs，Jimei University,Xiamen 361021,China)

摘要:: 基因组荧光原位杂交(GISH) 是分析远缘杂交染色体组结构的有力工具。为了提高杂交鲍GISH的信号强度，以皱纹盘鲍（Haliotis discus hannai）和杂色鲍（H.diversicolor）杂交子代为研究对象，研究了杂交缓冲液中探针质量浓度、封阻DNA质量浓度，以及去离子甲酰胺、硫酸葡聚糖或聚乙二醇6000的含量对GISH 信号强度的影响。结果表明，杂交缓冲液中，探针质量浓度应大于6.25 ng/μL，封阻DNA（鲑精DNA）的适宜质量浓度约为探针的10倍，去离子甲酰胺的适宜体积分数约为30%，硫酸葡聚糖的适宜体积分数为12.5%~17.5%。此外，用7.5%的聚乙二醇6000替代硫酸葡聚糖可以提高杂交信号，但存在信号噪点多等问题。

Abstract:: Genomic fluorescence in situ hybridization (GISH) is a powerful tool for analyzing the source of chromosomes in distant hybrid cells.In order to improve the signal of GISH，the effect of the probe dosage，blocked DNA concentration，deionized formamide concentration，and dextran sulfate or PEG6000 concentration on the signal intensity of GISH were investigated.The results showed that the appropriate concentration of the probe in the hybridization buffer was not less than 6.25 ng/μL；the concentration of the blocking DNA (salmon sperm DNA) should be 10 times the amount of the probe；and the suitable concentration of deionized formamide was about 30%，the suitable concentration of dextran sulfate is 12.5% to 17.5%.In addition，polyethylene glycol 6000 instead of dextran sulfate can increase the hybridization signal，with a suitable concentration of 7.5%.