A Streamlined Plant DNA Extraction Method with Liquid Nitrogen-Free Approach
Abstract
Molecular technique such as Polymerase Chain Reaction (PCR) is essential in various research fields. The amplification process of plant DNA can be challenging due to the presents of metabolites that can inhibit the polymerase eznyme, as well as expensive procedures or time-consuming laboratory work required. Most of the protocol involved liquid nitrogen, which is not always accessible, especially in laboratories with limited resources. Consequently, this study proposed an alternative protocol free from liquid-nitrogen usage, that was designed to be efficient in the DNA extraction from dry and fresh leaf samples across 40 plant species belonging to 27 different families. The DNA obtained from all the samples showed concentrations greater than 50 ng µL-1, with the quality indexes in the acceptable range (A260/280: 1.50-2.21, A260/230:0.60-2.20). The efficacy of this method was demonstrated by successful PCR amplification using rbcL primer, validating the DNA suitability. This protocol can be considered a good option to be used both with fresh and dried plant leaves. Moreover, the absence of liquid nitrogen usage in the protocol could decrease the laboratory cost considerably and turning it into a more easily replicable method to be used in laboratories with limited resources.
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