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Application of Anti-contamination in Gene Amplication Technique
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- Author:
- No author available
- Journal Title:
- JOURNAL OF CHINESE PHYSICIAN
- Issue:
- 12
- DOI:
- 10.3760/cma.j.issn.1008-1372.2004.12.004
- Key Word:
- PCR;UDG;微孔板杂交;抗污染
Abstract： 目的了解残留UDG对dU-DNA PCR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37℃放置20h杂交百分率;检测在-20℃、4℃、室温、37℃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13.281%～20.557%.t检验差异有显著性(t＝2.855,2.869,P＜0.05);经94℃灭活产物组37℃杂交A值比-20℃A值下降5.547%,未经94℃灭活杂交A值下降10.345%;94℃预变性30s杂交A值为98.714%、10min 30s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13.281%～20.557%与UDG有关.
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