Navidi W, Arnheim N, Waterman M S
Department of Mathematics, University of Southern California, Los Angeles 90089.
Am J Hum Genet. 1992 Feb;50(2):347-59.
A multiple-tubes procedure is described for using PCR to determine the genotype of a very small DNA sample. The procedure involves dividing the sample among several tubes, then amplifying and typing the contents of each tube separately. The results are analyzed by a statistical procedure which determines whether a genotype can be conclusively assigned to the DNA sample. Simulation studies show that this procedure usually gives correct results even when the number of double-stranded fragments in the sample is as small as 30. The procedure remains effective even in the presence of small amounts of laboratory contamination. We find that the multiple-tubes procedure is superior to the standard one-tube procedure, either when the sample is small or when laboratory contamination is a potential problem; and we recommend its use in these situations. Because the procedure is statistical, it allows the degree of certainty in the result to be quantified and may be useful in other PCR applications as well.
本文描述了一种用于通过聚合酶链式反应（PCR）确定极少量DNA样本基因型的多管程序。该程序包括将样本分至多个试管中，然后分别对每个试管中的内容物进行扩增和分型。通过统计程序分析结果，该程序可确定是否能明确将一种基因型归属于该DNA样本。模拟研究表明，即使样本中双链片段的数量少至30个，此程序通常也能得出正确结果。即使存在少量实验室污染，该程序仍然有效。我们发现，无论是样本量小还是存在实验室污染这一潜在问题时，多管程序均优于标准的单管程序；我们建议在这些情况下使用该程序。由于该程序具有统计性，它能对结果的确定程度进行量化，并且在其他PCR应用中可能也有用处。
