Development of Rapid and Less Hazardous Plant DNA Extraction Protocol using Potassium Phosphate Buffer.

Abstract

<b>Background and Objective:</b> Protocols commonly used in plant DNA extraction were known to be highly time-consuming and harmful due to the application of some hazardous reagents. Therefore, it was not applicable for such laboratories with limited resources as well as for high-throughput analysis. This study was aimed to develop a rapid yet less hazardous DNA extraction protocol for a plant using potassium phosphate buffer. <b>Materials and Methods:</b> Genomic DNA of chili pepper (<i>Capsicum annuum</i>) was extracted using potassium phosphate buffer and its efficacy was compared to three widely known protocols (CTAB-based, mini preparation and commercial kit). The extracted DNA from those four methods was evaluated based on its quality, quantity, practicality and cost per reaction. <b>Results:</b> Genomic DNA resulted from potassium phosphate buffer-based protocol exhibited comparable quality with adequate concentration for further downstream analysis. Results of PCR and sequencing were also emphasized the amplifiable DNA quality from this developed protocol. Compared to those commonly used protocols, potassium phosphate buffer consisted of 5 main working steps only, thus providing a simple yet rapid plant DNA extraction protocol. Since this protocol used ethanol only, it also offered a less hazardous and low-cost protocol that applicable for those resource-limited laboratories. <b>Conclusion:</b> This developed protocol provided a promising alternative of plant DNA extraction that might be applicable for both large scale analysis and any laboratory type. Further investigation was needed to evaluate its efficacy in extracting genomic DNA from various plants with different morphological characteristic.