Habibah, Noor Aini (2021) Artikel Keragaman Durian Berdasarkan Fragmen Internal Transcribed Spacers (ITS) DNA Ribosomal Melalui Analisis PCR-RFLP. Working Paper. Universitas Negeri Semarang.

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Abstract

Durian is one of the types of tropical fruits that has high economic value. Durian has an assortment of cultivars with morphology that are difficult to distinguish. Identity assurance is important for the fundamental information in increasing the efficiency of breeding and development of durian. Molecular identification is considered to be more accurate than the morphological characters. Polymerase Chain Reaction Restriction Fragment Length Polimorfism (PCR-RFLP) is a method to analyze the results of DNA fragment length difference that digest using restriction enzyme with endonuclease. The sample used in this study was the accession of durian taken randomly in Gunungpati district, Semarang. Genomic DNA was isolated based on Nucleon Phytopure Kit protocol with modification. ITS DNA ribosomal region was amplified using PCR-RFLP technique exploiting specific primers ITS L and ITS 4 produce ITS fragment length at 800 bp. Amplicon was digested using six restriction enzyme AluI, Eco471, Bsp1431, BsuRI, Mph11301 and Ade1. The research result of 11 accessions durian which is examined shows that the Bsp1431 enzyme has two specific sites that is cut on the size 550 bp and 120 bp. BsuR1 enzymes have a specific site cuts in the size of 600 bp. Whereas Eco471 enzyme has digesting sites specifics on size 450 bp. The conclusion of this research is durian accession number 12th, 15th, and 17th have close genetic relation and are alleged to be in one species.

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