Gel Purification of gDNA for Next-Generation Sequencing Applications
Supanut Utthiya, Passorn Wonnapinij, Pondpan Napaumpaiporn, Chokchai Kittiwongwattana, Jenjira Sakulkoo, Anongpat Suttangkakul, Supachai Vuttipongchaikij
Abstract
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.