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从随机扩增多态性DNA数据计算遗传相似系数:聚合酶链式反应产物的影响

Computing genetic similarity coefficients from RAPD data: the effects of PCR artifacts.

作者信息

Lamboy W F

机构信息

Department of Horticultural Sciences, Cornell University, Geneva, New York 14456-0462, USA.

出版信息

PCR Methods Appl. 1994 Aug;4(1):31-7. doi: 10.1101/gr.4.1.31.

Abstract

Random amplified polymorphic DNA (RAPD) markers have been used for many types of genetic analyses, including genome mapping, genotype fingerprinting, phylogeny reconstruction, and measuring genetic similarities. They suffer from one potential limitation, however, because the PCR that is used to produce informative amplification products often produces artifactual products as well. Optimization of PCR protocols to eliminate artifactual bands completely is often too costly or too time-consuming to be practical. Other methods for handling RAPD artifacts, such as deleting inconsistent or faint bands or using only those bands that are reproducible, introduce false negatives into the data. Simply ignoring artifacts and using all bands introduces false positives. When RAPD data are used to compute genetic similarity coefficients, such artifacts can cause significant bias in the estimation. The three coefficients most widely used with RAPD data, the simple matching coefficient, Jaccard's coefficient and Nei and Li's coefficient, differ in the amount of bias produced by a given level of artifactual bands. The simple matching coefficient and Nei and Li's coefficient always exhibit less percent bias than Jaccard's coefficient. For closely related organisms, Nei and Li's coefficient displays less percent bias than the simple matching coefficient. If new DNA samples possessing RAPD markers not present in the previously analyzed samples are added to a study, values of the simple matching coefficient will need to be computed for all samples, not just the new ones. Jaccard's and Nei and Li's coefficients, however, will not need to be recomputed. Furthermore, only Nei and Li's coefficient has a direct biological meaning (it is an estimate of the expected proportion of amplified fragments shared by two samples because they were inherited from a common ancestor). On the basis of these results, Nei and Li's coefficient is recommended for routine computation of genetic similarities using RAPD data, particularly if PCR artifacts are present.

摘要

随机扩增多态性DNA(RAPD)标记已用于多种类型的遗传分析,包括基因组作图、基因型指纹识别、系统发育重建以及测量遗传相似性。然而,它们存在一个潜在的局限性,因为用于产生信息性扩增产物的聚合酶链反应(PCR)通常也会产生人为产物。完全消除人为条带的PCR方案优化通常成本过高或耗时过长,不切实际。处理RAPD人为产物的其他方法,如删除不一致或模糊的条带或仅使用可重复的条带,会在数据中引入假阴性。简单地忽略人为产物并使用所有条带会引入假阳性。当使用RAPD数据计算遗传相似系数时,此类人为产物会在估计中导致显著偏差。与RAPD数据一起使用最广泛的三个系数,即简单匹配系数、杰卡德系数以及内氏和李氏系数,在给定水平的人为条带产生的偏差量上有所不同。简单匹配系数和内氏和李氏系数的偏差百分比总是比杰卡德系数小。对于亲缘关系密切的生物体,内氏和李氏系数的偏差百分比比简单匹配系数小。如果将具有先前分析样本中不存在的RAPD标记的新DNA样本添加到一项研究中,则需要为所有样本计算简单匹配系数的值,而不仅仅是新样本。然而,杰卡德系数和内氏和李氏系数不需要重新计算。此外,只有内氏和李氏系数具有直接的生物学意义(它是对两个样本由于继承自共同祖先而共享的扩增片段预期比例的估计)。基于这些结果,推荐使用内氏和李氏系数对RAPD数据进行遗传相似性的常规计算,特别是在存在PCR人为产物的情况下。

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