| 原位杂交中鲑鱼精DNA处理方法探究 |
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| 作者单位： | 山东省烟台市鲁东大学生命科学院，中国热带农业科学院热带生物技术研究所热带作物生物技术国家重点实验室，山东省烟台市鲁东大学生命科学院 烟台264025，海口，571101，烟台264025 |
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| 摘 要： | 采用机械打断/沸水浴和超声波处理两种方法对原位杂交中的封闭DNA——鲑鱼精DNA进行了断裂处理。分别探讨了不同的吹打次数、煮沸时间和超声波处理时间对鲑鱼精DNA片段大小的影响,并且对机械打断/沸水浴和超声波处理两种方法结果进行了比较分析。研究结果表明在煮沸时间20min时,吹打次数可根据所需原位杂交片段大小进行调整;超声波适宜打断时间为20min。
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| 关 键 词： | 鲑鱼精DNA 机械打断/沸水浴 超声波处理 |
Research on methods of salmon sperm DNA breaking in in situ hybridization |
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| Tingting Zhao,Jungang Wang,Jun Chen,Changliu Wang. Research on methods of salmon sperm DNA breaking in in situ hybridization[J]. Journal of Fujian Agriculture and Forestry University, 2007, 0(9) |
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| Authors: | Tingting Zhao Jungang Wang Jun Chen Changliu Wang |
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| Affiliation: | Tingting Zhao1,Jungang Wang2,Jun Chen1,Changliu Wang1* |
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| Abstract: | Two methods of mechanical break/boiling water bath and ultrasonic breaking were applied to break salmon sperm DNA which is usually used as blocking DNA in in situ hybridization. Different hypodermic syringe breaking times, boiling time and sonication period, relatative to salmon sperm DNA breaking segment size, were discussed and the efficiency of two methods was compared. The results show that when the boiling time is up to 20 minutes, special hypodermic syringe breaking times can be applied according to the using segment size in in situ hybridization and optimal treatment time is 20 minutes by ultrasonic breaking. |
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| Keywords: | Salmon sperm DNA mechanical break/boiling water bath ultrasonic breaking |
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