A Simple Solution Hybridization Rapidly Detects DNA by Fluorescent Probe

作者：毛强; 耿天龙; 王胜华

来源：光谱学与光谱分析, 2014, (6).

DOI：10.3964/j.issn.1000-0593(2014)06-1577-05

• Liquid hybridization
• FITC-1 2-dUTP
• Southern blot
• Specific DNA detection

Traditional Southern blot,as the golden standard to detect DNA,is widely used in molecular biolo-gy.However,its operation process has some significant disadvantages,such as radioactive contamination, complex procedures,time-consuming.Some new methods developed in recent years need expensive equipments and also have the defects of complex procedures and time-consuming.The paper described a method to detect DNA quickly by liquid hybridization.The probe used is a fragment of DNA sequence labeled by FITC-12-dUTP.The whole process includes four steps:probe preparation,hybridization,electrophoresis and signal de-tection.The comparative experiments using the single strand DNA probe and the double strand DNA probe in-dicated that the sensitivity of single strand DNA probes,which could detect 0. 38μg (1. 82 ×10-13 mol)plas-mid,is 2. 1 times than that of the double strand DNA probes which could detect 0. 8μg(3. 64×10-13 mol) plasmid.The total procedure is simple and can be completed in 3 h.

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