Ritzler M, Perschil I, Altwegg M
Department of Medical Microbiology, University of Zürich, Switzerland.
J Microbiol Methods. 1999 Feb;35(1):73-6. doi: 10.1016/s0167-7012(98)00104-3.
In diagnostic amplification protocols contamination by previously amplified nucleic acids is considered the major source of false positive results. Substituting dUTP for dTTP in the PCR and initial treatment with uracil-DNA glycosylase (UNG) virtually eliminates these carryover contaminations. Subsequent procedures to visualize the amplicons or to optimize sensitivity and specificity of the test are not always fully compatible with UNG-treated PCR products. Here we describe the more pronounced influence of residual UNG activity on the migration of PCR amplification products in polyacrylamide as compared to agarose gels.
在诊断扩增方案中，先前扩增的核酸污染被认为是假阳性结果的主要来源。在PCR中用dUTP替代dTTP并先用尿嘧啶-DNA糖基化酶（UNG）处理，实际上消除了这些残留污染。随后用于观察扩增子或优化检测灵敏度和特异性的程序并不总是与UNG处理的PCR产物完全兼容。在这里，我们描述了与琼脂糖凝胶相比，残留UNG活性对聚丙烯酰胺中PCR扩增产物迁移的更显著影响。
