Karakterisasi Molekuler Padi Transgenik dengan Beberapa Metode Isolasi DNA

Abstract

Rice is a major food commodity for agroindustrial country. Grain size is a major determinant of grain quality. One attempt that has been made in research to increase rice production is assembly of transgenic plants inserted with Os-GS3 (Oryza sativa Grain Size 3) gene. The material used in this study is cultivar Taipei 309. Rice plants that have been inserted with Os-GS3 gene were grown and the presence of the genes was analyzed through molecular characterization by PCR and Southern hybridization. DNA isolation methods used in this study were the simple method, miniprep scale, and large scale. They were compared to obtain the most efficient method in terms of turnaround time and resources used for PCR and Southern hybridization. PCR analysis of the three methods produced bands of the same size, in that 500 bp. Southern hybridization test of the DNA isolated using the large scale method produced bands or copy genes, where as that isolated using miniprep method did not produce band. The fast scale method is the most efficient DNA isolation method for PCR analysis, where as the large scale method is the most efficient DNA isolation method for Southern hybridization. The results in the silencing of gene expression of Os-GS3 with antisense RNA was successful when 4 transgenes integrated into the plant genome.