BioTechniques (Aug 2022)

Gel purification of gDNA for next-generation sequencing applications

  • Supanut Utthiya,
  • Passorn Wonnapinij,
  • Pondpan Napaumpaiporn,
  • Chokchai Kittiwongwattana,
  • Jenjira Sakulkoo,
  • Anongpat Suttangkakul,
  • Supachai Vuttipongchaikij

DOI
https://doi.org/10.2144/btn-2022-0013
Journal volume & issue
Vol. 73, no. 2
pp. 99 – 103

Abstract

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We demonstrate that gDNA can be conveniently and efficiently isolated and purified using standard agarose gel electrophoresis, band excision and gel purification. This method yields a substantial amount at microgram levels of gDNA per gel cleanup with high purity. An RNase A treatment step can be omitted. The quality of gDNA is suitable for next-generation sequencing, resulting in >10 Mb reads and high-quality read data (Phred score >28 up to 100 of 150 base reads). Furthermore, the gDNA can be kept intact in a gel slice for several days. This method has been tested for dictyostelids, bacteria and plants.

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