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基于白细胞计数评估DNA池构建的准确性。

Evaluating Accuracy of DNA Pool Construction Based on White Blood Cell Counts.

作者信息

Abrams Amy N, McDaneld Tara G, Keele John W, Chitko-McKown Carol G, Kuehn Larry A, Gonda Michael G

机构信息

Department of Animal Science, South Dakota State University, Brookings, SD, United States.

Department of Animal Science, Berry College, Mount Berry, GA, United States.

出版信息

Front Genet. 2021 Feb 5;12:635846. doi: 10.3389/fgene.2021.635846. eCollection 2021.

Abstract

Pooling individual samples prior to DNA extraction can mitigate the cost of DNA extraction and genotyping; however, these methods need to accurately generate equal representation of individuals within pools. The objective of this study was to determine accuracy of pool construction of blood samples based on white blood cell counts compared to two common DNA quantification methods. Fifty individual bovine blood samples were collected, and then pooled with all individuals represented in each pool. Pools were constructed with the target of equal representation of each individual animal based on number of white blood cells, spectrophotometric readings, spectrofluorometric readings, and whole blood volume with 9 pools per method and a total of 36 pools. Pools and individual samples that comprised the pools were genotyped using a commercially available genotyping array. ASReml was used to estimate variance components for individual animal contribution to pools. The correlation between animal contributions between two pools was estimated using bivariate analysis with starting values set to the result of a univariate analysis. Adonis test on distance matrix from the animal correlation showed clustering with method, and higher correlations between methods than within ( < 1 × 10). White blood cell count was predictive of sample representation when compared to pooling based on DNA concentration. Therefore, constructing pools using white blood cell counts prior to DNA extraction may reduce cost associated with DNA extraction and genotyping and improve representation of individuals in a pool.

摘要

在DNA提取之前合并个体样本可以降低DNA提取和基因分型的成本;然而,这些方法需要准确地在合并样本中生成个体的等量代表。本研究的目的是确定与两种常见的DNA定量方法相比,基于白细胞计数构建血液样本库的准确性。收集了50份个体牛血样本,然后将每个样本库中的所有个体合并。基于白细胞数量、分光光度读数、荧光分光光度读数和全血体积,以每个个体动物等量代表为目标构建样本库,每种方法有9个样本库,共36个样本库。使用市售的基因分型芯片对构成样本库的样本库和个体样本进行基因分型。使用ASReml估计个体动物对样本库贡献的方差成分。使用双变量分析估计两个样本库之间动物贡献的相关性,起始值设置为单变量分析的结果。对动物相关性距离矩阵进行的Adonis检验显示,样本库构建方法之间存在聚类,且方法之间的相关性高于方法内部(<1×10)。与基于DNA浓度的合并相比,白细胞计数可预测样本代表性。因此,在DNA提取之前基于白细胞计数构建样本库可能会降低与DNA提取和基因分型相关的成本,并改善样本库中个体的代表性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c8d/7893106/1ddd78946f42/fgene-12-635846-g001.jpg

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