OPTIMASI SUHU ANNEALING PRIMER DEGENERATE UNTUK MENGAMPLIFIKASI FRAGMEN GEN ARGININE DECARBOXYLASE (ADC) GENOM UBI KAYU LOKAL MALUKU TENGGARA

Abstract

Arginine decarboxylase (ADC) is an enzyme that plays a role in polyamine biosynthesis and has been shown to increase resistance to biotic and abiotic stress. Woody oak (Manihot esculenta Crantz) is known to grow and produce well in dry and poor nutrient conditions. The purpose of this study was to obtain optimum conditions in PCR reaction process to obtain candidate gene fragment of ADC. Four pairs of primers to amplify the gene fragments of ADC are degenerate from several plants that have been deposited on NCBI databases, namely Jatropa curcas (Acc XM_022220421), Populus trichocarpa (Acc XM_002306105.2), Capsicum annuum cv Nockwang (Acc KC160547.1) and Lycopersicon esculentum (Acc L16582.1). The success in amplifying a gene by PCR technique using a specially designed primer is determined by the precision of the primary attachment temperature with the DNA mold. Four primer pairs are designed to successfully amplify DNA sequence fragments from the local cassava genome from Malra, namely Malra012 and Malra016 genotypes. The MeadC1 primary pair can amplify the DNA mold and produce bands of less than 1,000 base pairs at a fixed temperature of 46 ° C.47 ° C. and 48 ° C.
Nucleotide base sequence analysis using primary pair MeadC1 has been done, but based on bioinformatic analysis using NCBI BLAST program, the obtained fragment did not show the encoding fragment of ADC gene.
Keywords : cassava, arginine decarboxylase, AADC