A Simple and Rapid DNA Isolation Method from Dry Pea Seeds Suitable for PCR Analyses

Abstract

Pea (Pisum sativum L.) is an important legume grown and consumed extensively worldwide. As a rich source of proteins, carbohydrates and vitamins, peas are important in human nutrition. In this study, we compared four different DNA isolation methods from four pea cultivars (F95-927, Specter, Windham and Nicoleta), with some modifications to the original extraction protocols: one SDS extraction buffer without PVP and one SDS extraction buffer with 1% PVP; one CTAB extraction buffer without PVP and one with 1% (1.5%) PVP. For all four extraction methods we used the same quantity of plant material (0.05 g) and an equal quantity of extraction buffer, so the comparison between methods to be very accurate. To establish which is the most efficient extraction method, after DNA isolation and purification, we submitted our samples to PCR analyzes with two markers: ISSR marker 17899A and SSR marker AA175. In my study, based on spectrophotometric measurements and PCR results, I concluded that both CTAB extraction methods were not suitable for DNA extraction from dry pea seeds because they did not show amplification products. The most appropriate DNA extraction method was SDS1 which provided a good quality DNA.