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Genotyping of Plant and Animal Samples without Prior DNA Purification
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Direct single-nucleotide polymorphism genotyping from whole blood without DNA extraction.

Risako Furukawa1, Teruaki Tozaki1, Koki Kawate1

  • 1Genetic Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi 320-0851, Japan.

Journal of Equine Science
|March 20, 2026
PubMed
Summary

Researchers developed a new method for direct single-nucleotide polymorphism (SNP) genotyping from whole blood, eliminating the need for DNA extraction. This technique offers a simple, cost-effective approach for genetic analysis in various fields.

Keywords:
LCORLMSTNhorsesingle-nucleotide polymorphism genotype

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Area of Science:

  • Genetics
  • Molecular Biology
  • Biotechnology

Background:

  • Single-nucleotide polymorphisms (SNPs) are crucial genetic markers associated with diverse phenotypes.
  • Traditional SNP genotyping requires DNA extraction, a time-consuming and costly process.
  • Efficient SNP genotyping is vital for medical, veterinary, and performance-related genetic studies.

Purpose of the Study:

  • To develop and validate a novel, direct SNP genotyping method from whole blood.
  • To eliminate the requirement for DNA extraction in SNP analysis.
  • To assess the method's simplicity, cost-effectiveness, and versatility for genetic targets.

Main Methods:

  • Whole blood samples were diluted and analyzed using real-time polymerase chain reaction (PCR) with hydrolysis probes.
  • Optimization involved increasing annealing temperature and PCR cycle number to enhance specificity and sensitivity.
  • Genotyping was performed for SNPs in *MSTN* and *LCORL* genes.

Main Results:

  • The novel method achieved direct SNP genotyping from whole blood without prior DNA extraction.
  • Results demonstrated complete concordance with conventional real-time PCR methods using extracted DNA.
  • The technique proved effective for SNPs in *MSTN* and *LCORL*, with potential for other targets like *CDH13* and *MAOA*.

Conclusions:

  • A simple, low-cost, and versatile direct SNP genotyping method from whole blood was successfully developed.
  • This approach bypasses DNA extraction, streamlining genetic analysis.
  • The method shows broad applicability for various genetic targets in research and diagnostics.