Optimized double-digest genotyping by sequencing (ddGBS) method with high-density SNP markers and high genotyping accuracy for chickens

PLoS One. 2017 Jun 9;12(6):e0179073. doi: 10.1371/journal.pone.0179073. eCollection 2017.

Abstract

High-density single nucleotide polymorphism (SNP) markers are crucial to improve the resolution and accuracy of genome-wide association study (GWAS) and genomic selection (GS). Numerous approaches, including whole genome sequencing, genome sampling sequencing, and SNP chips are able to discover or genotype markers at different densities and costs. Achieving an optimal balance between sequencing resolution and budgets, especially in large-scale population genetics research, constitutes a major challenge. Here, we performed improved double-enzyme digestion genotyping by sequencing (ddGBS) on chicken. We evaluated eight double-enzyme digestion combinations, and EcoR I- Mse I was chosen as the optimal combination for the chicken genome. We firstly proposed that two parameters, optimal read-count point (ORP) and saturated read-count point (SRP), could be utilized to determine the optimal sequencing volume. A total of 291,772 high-density SNPs from 824 animals were identified. By validation using the SNP chip, we found that the consistency between ddGBS data and the SNP chip is over 99%. The approach that we developed in chickens, which is high-quality, high-density, cost-effective (300 K, $30/sample), and time-saving (within 48 h), will have broad applications in animal breeding programs.

MeSH terms

  • Animals
  • Chickens / genetics*
  • Computational Biology / methods
  • Genotype
  • Genotyping Techniques*
  • High-Throughput Nucleotide Sequencing*
  • Polymorphism, Single Nucleotide*
  • Reproducibility of Results

Grants and funding

This study was financially supported by grants from the 948 Program of the Ministry of Agriculture of China (2012-G1[4]), the National High Technology Research and Development Program of China (2013AA102501), and the National Natural Science Foundation of China (NSFC, 31272432).