A simplified protocol for the recovery of high quality DNA from nutmeg

P. Diyasree, K.A. Lokesh, P. Lidiya John, Tintu Joy, V. Teena Ramachandran, Anu Thomas Muttakulath, T.M. Manila, V.T.K. Sandhya Rajan, Deepu Mathew

Abstract


High levels of secondary metabolites even in the immature tissues leave the process of quality DNA isolation very difficult in the genus Mysristica. This difficulty is often characterized in the form of lower levels of DNA recovery and poor absorbance ratios due to RNA and protein contaminations, while analyzed in a spectrophotometer. The quantity and quality of DNA obtained through different methods were comparatively evaluated and the conditions for suitable recovery were optimized. A CTAB based simple and reliable method, standardized with necessary modifications on the protocol given by Doyle and Doyle was found to yield the best in terms of DNA quantity and quality. This method when compared with other protocols such as Rogers and Bendich and the Murray and Thompson, which are reported by the previous workers, was proven superior. The DNA recovery from the tender leaf tissues was high in the mean range of 83.55 μg/g with a purity of 1.79. The standardized protocol was further experimented in other phenolic-rich plants such as cashew and cocoa and was found to yield high quality DNA.


Keywords


DNA isolation; Extraction; Genomics; Molecular markers; Myristica fragrans

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A KAU publication [CODEN: JTAGEI; ISSN 0971-636X; eISSN 0973-5399]