GISH中杂交鲷亲本封阻DNA不同制备方法的效果

Effect of Different Preparation Methods by Hybrid Porgy's Blocking DNA in Gish

目的:基因组荧光原位杂交(GISH)作为一种细胞遗传学的研究方法,以其特定的优点被越来越广泛的应用于杂种染色体分析、染色体行为考察等方面。采用适当的封阻DNA制备方法得到合适的封阻DNA从而使得在基因组原位杂交中大大提高杂交效率,更能突显GISH方法的优势。方法:采用煮沸法和超声波破碎法对杂交鲷亲本真鲷(♀)、黑鲷(♂)DNA进行剪切研究了两种鲷类封阻DNA的制备。结果:煮沸15 min时DNA已被切断。煮沸70 min以后,DNA片段更加集中,主要分布在500bp左右。煮沸90 min或者煮沸120 min以后发现,DNA电泳条带主要分布于250 bp附近,已达到GISH所需封阻DNA片段大小的要求。结论:煮沸法对基因组DNA的剪切,与超声波清洗仪剪切DNA相比,具有更高的效率。研究结果为基因组荧光原位杂交的顺利进行提供了条件。

Objective:Genomic in situ hybridization(GISH) as a method study of cytogenetic with the specific advantage was extensively used in the hybrid-Chromosome analysis and chromosome behavior canvass.Proper block can greatly promote the efficiency of gene recombination.Method:Pagrosomus major and Sparus macrocephalus genomic DNA was cut by boiling and ultrasonic methods to study the preparation of blocking DNA.Result:The results showed that the efficiency of boiling method was higher than ultrasonic cutting method,and boiling method was more easily operated.Boiling for 15 min,the DNA fragment was turn to be cut.For 70 min,the fragments were more Concentration,most DNA length was about 500 bp.Conclusion:When genomic DNA was boiled for 90 min-120 min,he fragments were concentrated in 250 bp,thus DNA was suitable for blocking DNA in GISH.These findings laid a foundation for further study on GISH.