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目的 了解残留UDG对dU -DNAPCR产物的影响及灭活UDG时对Taq -P的影响。方法 采用微孔板杂交技术检测dU -DNAPCR产物 3 7℃放置 2 0h杂交百分率 ;检测在 -2 0℃、4℃、室温、3 7℃时不同保存条件下的杂交百分率 ;94℃灭活 10min对Taq -P活性的影响。结果 加 0 2单位、1 0单位UDG组比不加UDG组杂交A值下降 13 2 81%～ 2 0 5 5 7%。t检验差异有显著性 (t=2 85 5 ,2 869,P <0 0 5 ) ;经 94℃灭活产物组 3 7℃杂交A值比 -2 0℃A值下降 5 5 47% ,未经 94℃灭活杂交A值下降 10 3 45 % ;94℃预变性 3 0s杂交A值为 98 714 %、10min 3 0s组杂交A值为 96 818%。结论 以上结果提示经 94℃加热灭活对PCR产物保存有重要作用 ,不仅对灭活UDG有作用而且还可灭活来源于UDG以外可破坏DNA的酶。表明含UDG组杂交A值比不加UDG组低13 2 81%～ 2 0 5 5 7%与UDG有关。
Objective To investigate the effects of residual UDG on dU-DNA PCR products and the effect of UDG on Taq-P. Methods The hybridization rate of dU-DNA PCR products at 37 ℃ for 20 h was detected by microplate hybridization. The hybridization percentages at different storage conditions at -20 ℃, 4 ℃, room temperature and 37 ℃ were determined. After inactivation at 94 ℃ for 10min Effect on Taq-P activity. Results With 0 2 units, the A value of 10 UDG group decreased 13 2 81% -2 205 7% compared with that of the UDG group. (t = 2 855, 2 869, P <0 05). The value of A at 37 ℃ after inactivation at 94 ℃ decreased by 55 47% compared with that at -2 0 ℃ After inactivation, the value of A was decreased by 10 3 45% after inactivation at 94 ° C. The A value of hybridization at 94 ° C for 30 s was 98 714%, and the value of hybridization A was 96 818% at 10 min 3 s. Conclusion The above results suggest that heat inactivation at 94 ℃ plays an important role in the preservation of PCR products, not only inactivating UDG but also inactivating enzymes that can destroy DNA other than UDG. The results showed that the A value of hybridization with UDG group was lower than that of UDG group by 13.281% -2507% and UDG.