Study on the Effect of Residual Uracil DNA Glycosylase upon HCV cDNA
- VernacularTitle:残留尿苷酶对HCV cDNA破坏作用的研究
- Author:
Shaocai DU
;
Ying JI
;
Ling ZHU
- Publication Type:Journal Article
- Keywords:
Uracil DNA glycosylase(UDG);
PCR;
dUTP4
- From:
Journal of Chinese Physician
2000;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
To explore the destructive effect of residual uracil DNA glycosylase on dUTP incorporation to the cDNA PCR products,PAGE and DNA-EIA hybridization technique were applied to detect the efficiency of anti-contamination and the rate of DNA hybridization.In this study,it was found that:⑴Good effect could be achieved with the reaction of 2u of uracil DNA glycosylase(UDG)for 20min at 37℃.⑵UDG could be inactivated at 94℃ for 10min.The hybridization rate of control group of PCR products was 94 18% at room temperature for 65 hours.The hybridization rates of test groups at-20℃ for 65 hours were 87 69%,77 24%,76 83%,respectively.And the inactivation times of anti-contamination groups at 94℃ were“0”min,“10”min and “20”min,respectively;at room temperature for 65 hours,these hybridization rates were 74 77%,72 50%,70 29%,respectively.There were significant differences between control and rest groups(P0 05).Our study suggested that the hybridization rate of test groups kept at-20℃ decreased 19 41%,after 65 hours,the rate decreased 27 45%,but in control group only decreased 5 82% for 65h at room temperature.So that,we conclude that the residual UDP can cause above results.
