RHIZOSPHERE SOIL SAMPLING METHOD OF BANANA PLANT FOR DNA EXTRACTION

Abstract

This invention relates to a method for sampling banana plant rhizosphere soil for DNA extraction used in microbiome analysis using NGS, particularly for sampling in locations where laboratory facilities are not available. The process of sampling rhizosphere soil until the sample is ready for the DNA extraction stage is a very important step to ensure the quality of the next Next Generation Sequencing (NGS) results. Sampling of banana plant rhizosphere soil to obtain quality DNA that can be used in microbiome analysis is carried out using a method consisting of a combination of the use of Phosphate Buffer Saline (PBS) solution to dissolve the rhizosphere soil, a sedimentation and incubation process at a temperature of 4 - 8 oC for 8 - 12 hours to produce rhizosphere soil pellets, and the use of DNA / RNA preservation solutions to maintain DNA stability.The DNA extraction results from the sample were of high quality with a concentration of more than 10 nanograms / microliter (ng / µL) and purity at an A260 / 280 ratio of 1.94 – 2.13. The results of PCR amplification and visualization by agarose gel electrophoresis showed that the banana rhizosphere soil DNA successfully amplified the target hypervariable region of 16S rRNA V3-V4 with the appropriate size of ~550 bp. This method is a sampling method in locations where laboratory facilities are not available.