Pengembangan Metode Isolasi Total RNA, mRNA dan Sintesis cDNA Sengon (Falcataria moluccana L. Nielsen) yang Sehat dan Terserang Hama Boktor (Xystrocera Festiva).

Abstract

Sengon (Falcataria moluccana L. Nielsen) has a natural self-defense mechanism towards pest infection by synthesizing certain macromolecul such as tripsin inhibitor and alfa-amilase inhibitor. Characterization of those genes which produce the inhibitor need to be done on resistant trees and the vulnerable ones, to define the differences so that can be used for tree improvement. This research aim to develop an appropriate isolation methods of total RNA, mRNA and cDNA of plants that have a inhibitor enzymes such as sengon. The materials used in this research were cambium and timber tissue from resistant and vulnerable tree. Total RNA was isolated using Total RNA Mini Kit by Geneaid, mRNA was isolated using PolyATract mRNA Isolation System III with Magnetic Stand by Promega and cDNA synthesize was conducted using cDNA Substraction Kit by Clontech and RevertAid RT Transcription Kit by Thermo. The highest concentration of total RNA isolation were 65,60ng/μl from modification methods, and the best mRNA yields was resulted by modification methods. The highest concentration for cDNA synthesize were 2296,17ng/μl from modification methods.