Study on the Effect of Residual Uracil DNA Glycosylase upon HCV cDNA / 中国医师杂志

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 that⑴Good 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(P0 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.