Capillary electrophoresis and laser-induced fluorescence (CE-LIF) for screening of wild type genes based on molecular beacons hybridization
- VernacularTitle:分子信标与毛细管电泳——激光诱导荧光检测联合筛查野生型基因
- Author:
Xiaofeng JIANG
;
Jianing CHEN
;
Fuzhen LV
;
Jinhui WANG
;
Hongyan LIANG
;
- Publication Type:Journal Article
- Keywords:
Molecular beacons;
Electrophoresis, capillary;
Gene;
Polymorphism, single strand conformational
- From:
Chinese Journal of Laboratory Medicine
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objectives A new method for the detection of PCR ampli fied DNA in capillary electrophoresis and laser induced fluorescence (CE LIF) system based on molecular beacons hybridization is described.Methods The method is based on the hybridization of molecular beacons into the amplification product, then separating the hybridized product with CE LIF system. This method was applied to the detection of p73exon3, p53exon5 and p16exon2 for gastric carcinoma. Results Detected with CE LIF molecular beacons method, the wild type percent of p73exon3 is 100%, same as SSCP; p53exon5 is 60 0%, while SSCP is 63 3%, P=0 99; p16exon2 is 20 0%, same as SSCP(20 0%).Conclusion The results demonstrate that the specific peaks appeared in wild type samples while no peaks or unspecific peaks in mutation ones. The results showed correlation with the results of PCR SSCP. The method is able to develope as a clinical screening method for tumor suppressor genes to diagnose carcinoma.
