Zimmermann K, Schwarz H P, Turecek P L
Baxter Hyland Immuno, Vienna, Austria.
Comp Med. 2000 Jun;50(3):314-6.
In an attempt to find a rapid and reproducible method for routine polymerase chain reaction genotyping of transgenic mice, two novel approaches were developed.
One approach allowed reproducible amplification from crude lysates of tail snips, using a thermally activated polymerase. In a second approach, for situations in which non-invasive techniques are necessary, oral swab specimens were amplified after DNA extraction by use of an isolation kit. Samples from 10 transgenic factor VIII knockout mice were genotyped after processing by use of these and other methods.
False-negative results were not obtained by use of the two novel approaches. Despite their relative simplicity, both approaches yielded results comparable to those obtained by use of procedures known to be reliable, such as organic extraction and a DNA extraction kit.
Both approaches are useful for PCR-amplification of DNA from mammalian sources.
为了找到一种快速且可重复的转基因小鼠常规聚合酶链反应基因分型方法，开发了两种新方法。
一种方法使用热激活聚合酶，可从尾部剪下物的粗裂解物中进行可重复的扩增。在第二种方法中，对于需要非侵入性技术的情况，使用分离试剂盒提取DNA后，对口腔拭子标本进行扩增。使用这些方法和其他方法处理来自10只转基因因子VIII基因敲除小鼠的样本后进行基因分型。
使用这两种新方法未获得假阴性结果。尽管它们相对简单，但两种方法产生的结果与使用已知可靠的程序（如有机提取和DNA提取试剂盒）获得的结果相当。
这两种方法都可用于从哺乳动物来源的DNA进行PCR扩增。
