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中国农学通报 ›› 2018, Vol. 34 ›› Issue (17): 32-38.doi: 10.11924/j.issn.1000-6850.casb17080020

• 生物技术科学 • 上一篇    下一篇

核酸染料Gelred在两种凝胶检测系统中的应用研究

余虹莹,翟悦,吴则东   

  1. 黑龙江大学生命科学院,黑龙江大学生命科学院,黑龙江省普通高校甜菜遗传育种重点实验室/黑龙江大学
  • 收稿日期:2017-08-04 修回日期:2017-08-29 接受日期:2017-08-31 出版日期:2018-06-20 发布日期:2018-06-20
  • 通讯作者: 吴则东
  • 基金资助:
    国家糖料现代农业产业技术体系甜菜高品质品种改良岗位专项基金(CARS-170111)。

The Application of Nucleic Acid Dye Gelred in Two Gel Detection Systems

  • Received:2017-08-04 Revised:2017-08-29 Accepted:2017-08-31 Online:2018-06-20 Published:2018-06-20

摘要: 由于无毒的核酸染料Gelred价格比较昂贵,在不影响实验效果的情况下,探讨Gelred在聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳中的最小用量。笔者将10000×的核酸染料Gelred稀释成1×、10×、20×、30×、40×、50×,将稀释后的核酸染料直接加入到甜菜PCR产物中。将核酸染料和甜菜PCR产物的混合物直接点样聚丙烯酰胺凝胶和琼脂糖凝胶,电泳后直接利用凝胶成像系统进行检测。以及另外一种方法是利用1×的Gelred分别对聚丙烯酰胺凝胶和琼脂糖凝胶进行泡胶,用凝胶成像系统进行检测。结果表明,Gelred 30×及以上的稀释浓度均适合于聚丙烯酰胺凝胶和琼脂糖凝胶,而1×的Gelred稀释液更适合于对聚丙烯酰胺凝胶进行泡胶。将稀释后的Gelred直接和PCR产物混合后,配合聚丙烯酰胺凝胶电泳,不仅减少了Gelred的使用量,而且减少了显影的时间。

关键词: ‘阿巴特’梨, ‘阿巴特’梨, 采后处理, 冷藏, 货架期, 品质

Abstract: Because nontoxic nucleic acid dye Gelred has high cost, the minimum dosage of Gelred is investigated on non-denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis, without affecting the experimental results. 10000×Gelred was individually diluted to 1×, 10×, 20×, 30×, 40× and 50×, directly mixed with the PCR production of sugar beet, and then the mixture was directly detected by nondenaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis. The detection was conducted with GEL imaging system. The other method was that the non-denaturing polyacrylamide gel and agarose gel were soaked in 1×Gelred and then showed by GEL imaging system. The results showed that 30×Gelred and above-30×Gelred were suitable for the detection of non-denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis, and 1×Gelred was more suitable for the soak of non-denaturing polyacrylamide gel. The dilute Gelred was mixed with PCR production, and then coordinated with non- denaturing polyacrylamide gel electrophoresis, which not only reduced the amount of Gelred, but also decreased the developing time.