Developing and Testing Specific Primers for PHYA Gene Sequence Analysis in Fourteen Indonesian sorghum varieties Siti Halimah larekeng, Firzan Nainu, Muh. Restu, Ahmad Rifqi Makkasau, Nasri Nasri, Yuni Fitri Cahyaningsih
1. Natural heritage and Biodiversity Research Center Hasanuddin University, Makassar
2. Faculty of Pharmacy, Hasanuddin University, Makassar
3. Faculty of Vocational , Hasanuddin University, Makassar
4. Faculty of Forestry, Hasanuddin University, Makassar
5. National Research and Innovation Agency, Indonesia
Abstract
Primer is one of the important components needed in the DNA synthesis process using a PCR machine. A specific primer can be developed to amplify the target gene by utilizing Primer3Plus and Python. This study developed specific primers to amplify the PHYA gene using Primer3Plus and compared its amplification rate with primers developed using Python. Sequence Sorghum bicolor phytochrome A (PHYA) gene, (GenBank: AY466073.1) is used as a template to generate the primary sequence. Both methods produced four pairs of primers to amplify the PHYA gene. Both methods were able to produce primers that could amplify the gene in fourteen varieties of sorghum. Each primer tested produces a clear band. The optimum temperature ranges from 51.5 C-57.5 C with PCR-sized products around 950bp. The internet network is needed to be able to design primaries using Primer3Plus while Python can be used without an internet connection but requires the ability to understand Python.
Keywords: Clear bands, DNA amplify, Primers, Specific gene
