Simple, fast and inexpensive hot sodium hydroxide and tris DNA extraction method for genotyping tomato and melon seeds

Biotechniques. 2023 Dec;75(6):245-249. doi: 10.2144/btn-2023-0054. Epub 2023 Nov 2.

Abstract

Seed commerce is a highly profitable global market. Most commercialized seeds are hybrid seeds originating from a controlled cross between two selected parental lines. The market value of hybrid seeds depends on their hybrid genetic purity. DNA molecular markers are a reliable and widespread tool to genotype plant materials; however, DNA extraction from seeds is challenging, often laborious and expensive. With the ultimate goal of creating a tomato and melon hybrid seeds purity test, various challenges arise. To overcome these problems and with the purpose of crude DNA extraction, a simple, fast, inexpensive and easily scalable adaptation of the hot sodium hydroxide and tris method coupled to a competitive allele-specific PCR genotyping method is proposed.

Keywords: Cucumis melo; KASP; Solanum lycopersicum; crude DNA extraction; hybrid seed; molecular markers; seed purity test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cucurbitaceae* / genetics
  • DNA
  • Genotype
  • Seeds / genetics
  • Sodium Hydroxide
  • Solanum lycopersicum* / genetics

Substances

  • Sodium Hydroxide
  • DNA