核酸染料Gelred在两种凝胶检测系统中的应用研究

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

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×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,然后点样到聚丙烯酰胺凝胶和琼脂糖凝胶上,电泳后直接利用凝胶成像系统进行检测。另外一种方法是利用1×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×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。

Because nontoxic nucleic acid dye Gelred has high cost,the minimum dosage of Gelred isinvestigated on non-denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis,withoutaffecting 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 non-denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis.The detection was conductedwith GEL imaging system.The other method was that the non-denaturing polyacrylamide gel and agarose gelwere soaked in 1×Gelred and then showed by GEL imaging system.The results showed that 30×Gelred andabove-30×Gelred were suitable for the detection of non-denaturing polyacrylamide gel electrophoresis andagarose gel electrophoresis,and 1×Gelred was more suitable for the soak of non-denaturing polyacrylamidegel.The dilute Gelred was mixed with PCR production,and then coordinated with non-denaturingpolyacrylamide gel electrophoresis,which not only reduced the amount of Gelred,but also decreased thedeveloping time.