Low-cost methods for DNA extraction and quantification

Abstract

Nucleotide variation, whether induced or natural, is responsible for a vast majority of heritable phenotypic variation. Evaluation of genomic DNA sequence is therefore fundamental for both functional genomics and markerassisted breeding. The starting point for this is the extraction of high-quality DNA of a suitable quantity for planned downstream applications. A myriad of kits and techniques exists, but not all are suitable for laboratories with limiting infrastructure due to high costs and reliance on toxic chemicals. We describe here a protocol for extraction of high-quality genomic DNA from leaves of a variety of plant species that uses self-prepared buffers and reagents and obviates the need for toxic organic chemicals such as chloroform. We also provide a protocol for using free image analysis software to quantify isolated DNA.