Isolation and Characterization of DNA from Root Tubers

Abstract

This study reports a modified SDS extraction technique for isolation of DNA from root tubers namely Dioscorea (yam), Manihot esculenta (cassava) and Colocasia esculenta (cocoyam). DNA extraction in many plants is very difficult because of secondary metabolites that interfere with DNA isolation procedure. Young fresh leaves were collected from the selected tubers and DNA extracted using SDS-extraction protocol, the DNA isolated from the extracted leaves was subjected to gel electrophoresis for determining their purity and concentration and PCR amplification was carried out on the extracted DNA, afterwards gel-electrophoresis was performed for molecular weight determination of the samples. DNA sequencing were performed on both reverse and forward using Sanger sequencing method which gave the nucleotide bases of the samples. DNA isolated by modified SDS protocol method was pure and highest level of purity was obtained from Manihot esculenta, Colocasia esculenta and Dioscorea which are 1.77, 1.72 and 1.20 with concentration of 537.7 ng/µl, 992.4 ng/µl and 149.8 ng/µl respectively. The DNA isolated from root tubers was pure and of good quality, and the isolated DNA was characterized by sequencing. The isolated DNA was successfully sequenced which coded for marker genes of the respective root tubers.