SNP detection for massively parallel whole-genome resequencing

Genome Res. 2009 Jun;19(6):1124-32. doi: 10.1101/gr.088013.108. Epub 2009 May 6.

Abstract

Next-generation massively parallel sequencing technologies provide ultrahigh throughput at two orders of magnitude lower unit cost than capillary Sanger sequencing technology. One of the key applications of next-generation sequencing is studying genetic variation between individuals using whole-genome or target region resequencing. Here, we have developed a consensus-calling and SNP-detection method for sequencing-by-synthesis Illumina Genome Analyzer technology. We designed this method by carefully considering the data quality, alignment, and experimental errors common to this technology. All of this information was integrated into a single quality score for each base under Bayesian theory to measure the accuracy of consensus calling. We tested this methodology using a large-scale human resequencing data set of 36x coverage and assembled a high-quality nonrepetitive consensus sequence for 92.25% of the diploid autosomes and 88.07% of the haploid X chromosome. Comparison of the consensus sequence with Illumina human 1M BeadChip genotyped alleles from the same DNA sample showed that 98.6% of the 37,933 genotyped alleles on the X chromosome and 98% of 999,981 genotyped alleles on autosomes were covered at 99.97% and 99.84% consistency, respectively. At a low sequencing depth, we used prior probability of dbSNP alleles and were able to improve coverage of the dbSNP sites significantly as compared to that obtained using a nonimputation model. Our analyses demonstrate that our method has a very low false call rate at any sequencing depth and excellent genome coverage at a high sequencing depth.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Asian People / genetics
  • Chromosomes, Human, X / genetics
  • Computational Biology / methods
  • Genetics, Population / methods
  • Genome, Human / genetics*
  • Genotype
  • Humans
  • Likelihood Functions
  • Models, Genetic
  • Models, Statistical
  • Polymorphism, Single Nucleotide*
  • Probability
  • Reproducibility of Results
  • Sequence Analysis, DNA / instrumentation
  • Sequence Analysis, DNA / methods*
  • Software*