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

Autor:	Abrams AN; Department of Animal Science, South Dakota State University, Brookings, SD, United States.; Department of Animal Science, Berry College, Mount Berry, GA, United States., McDaneld TG; Genetics, Breeding, and Animal Health Research Unit, U.S. Meat Animal Research Center, USDA-ARS, Clay Center, NE, United States., Keele JW; Genetics, Breeding, and Animal Health Research Unit, U.S. Meat Animal Research Center, USDA-ARS, Clay Center, NE, United States., Chitko-McKown CG; Animal Health Genomics Research Unit, U.S. Meat Animal Research Center, USDA-ARS, Clay Center, NE, United States., Kuehn LA; Genetics, Breeding, and Animal Health Research Unit, U.S. Meat Animal Research Center, USDA-ARS, Clay Center, NE, United States., Gonda MG; Department of Animal Science, South Dakota State University, Brookings, SD, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in genetics [Front Genet] 2021 Feb 05; Vol. 12, pp. 635846. Date of Electronic Publication: 2021 Feb 05 (Print Publication: 2021).
DOI:	10.3389/fgene.2021.635846
Abstrakt:	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 ( P < 1 × 10 -6 ). 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. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2021 Abrams, McDaneld, Keele, Chitko-McKown, Kuehn and Gonda.)
Databáze:	MEDLINE
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