<span style="font-size:11.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB">Optimization of DNA isolation protocol of <i style="mso-bidi-font-style: normal">Aconitum balfourii</i> Stapf.: A rare medicinal herb of Himalayan alpine</span>

Abstract

<i style="mso-bidi-font-style:normal">Aconitum balfourii</i> Stapf. (Family: Ranunculaceae) is a rare medicinal herb distributed at high altitude (2800-4200 m) of Himalayan region. It is rich in secondary metabolites, especially aconitines, having medicinal properties. Secondary metabolites, mainly polyphenols, cause hinderance in the isolation of good quality DNA from the plant. Therefore, in the present investigation, an attempt was made to determine the total phenolic contents in various parts of <i style="mso-bidi-font-style:normal">in vitro</i> grown plantlets and DNA isolation was optimized from<i style="mso-bidi-font-style: normal"> in vitro</i> grown fresh leaves. In the process, 2.5% polyvinyl pyrrolidone was found suitable for obtaining phenolic free DNA. The method also used high salt concentration (5 M NaCl) to remove polysaccharide. In the optimized protocol, DNA isolated with or without liquid nitrogen was comparable in terms of yield and quality. Thus, the suggested protocol provides the contaminant free DNA with good yield that can be used in various molecular biology applications.