摘要
在聚丙烯酰胺凝胶法检测DNA过程中,核酸染料一旦与DNA结合其荧光量子急剧增加,在紫外光或者可见光的照射下就可以观测到DNA。但目前尚无核酸染料对聚丙烯酰胺凝胶中DNA不同染色方法的比较研究。本研究比较了UltraPower^(TM)和G-Red两种无毒核酸染料对聚丙烯酰胺凝胶法检测DNA的四种染色方法(点染,胶染,点染+胶染,泡染),结果表明:UltraPowerTM和G-Red两种核酸染料通过泡染法成功检测到凝胶中的大小片段DNA,条带清晰易辨;DNA检测灵敏性高,都在80 pg以下;染色过程安全简单,只需要一种无毒核酸染料和一个染色步骤。但UltraPower^(TM)检测DNA条带的灵敏性(19.53 pg)是G-Red的4倍(78.13 pg),而每块胶DNA染色成本(2.65元)仅是后者的1/2 (5.30元)。从经济和DNA检测灵敏性两方面考虑,UltraPower^(TM)染色比G-Red更胜一筹。核酸染料泡染法将促进聚丙烯酰胺凝胶中DNA的高效灵敏检测。
The property of DNA can be visible under ultraviolet or white lights when nucleic acid dyes bind to DNA to induce their fluorescent quantum yield to enhance dramatically,which makes them suitable for polyacrylamide gel DNA detection.However,little is known about the efficiency of different polyacrylamide gel DNA staining ways for these dyes.In this study,two nucleic acid dyes of UltraPower^(TM) and G-Red were used to detect DNA bands in polyacrylamide gels with four different DNA stain ways:DNA staining,gel-made staining,DNA+gel-made staining,and postelectrophoretic staining.Results show that for these two dyes,a post-electrophoresis stain significantly outstrips the other three staining ways,clearly detecting both small and long DNA fragments;exhibiting a less than 80 pg sen sitivity requiring only one reagent and one staining step to finish the whole stain process.However,UltrapowerM sensitivity(19.53 pg)and cost(￥2.65)per stain gel is fourfold and a half that ofG-Red's(78.13 pg and ￥5.30).Ultrapower^(TM) stain performs more excellent than G-Red stain in consideration of its sensitivity and economy.The postelectrophoresis stain of nucleic acid dyes will facilitate the efficient and sensitive DNA polyacrylamide gel detection.
作者
缪迎春
李瑾
刘子睿
高成杰
崔凯
Miao Yingchun;Li Jin;Liu Zirui;Gao Chengjie;Cui Kai(Institute of Highland Forest Science,Chinese Academy of Forestry,Kunming,650233)
出处
《分子植物育种》
CAS
北大核心
2023年第8期2635-2641,共7页
Molecular Plant Breeding
基金
中央级公益性科研院所基本科研业务费专项(No.CAFYBB2019ZB006)
云南省基础研究计划重点项目(No.2019FA013)共同资助。
