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| Abstract : Perpustakaan Tuanku Bainun |
| This study is to establish a Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method and diagnostic PCR based on nuclear DNA internal transcribed spacer 2 (ITS2) for E. longifolia, L. pumila, and O. stamineus. PCRRFLP was then used to validate the species identification for herbal products. The PCRRFLP was developed for rapid identification using restriction enzymes TaqI, BamHI, HinfI, EcoRI, EcoRV, Mbol, and Mspl. The ITS2 sequences were identified and compared between plant specimens of E. longifolia, L. pumila, and O. stamineus with 106 samples of commercial herbal products. As a result, plant specimens of E. longifolia, L. pumila, and O. stamineus were successfully identified with high similarity of 100%, 100%, and 99.33%, respectively, based on the National Center for Biotechnology Information (NCBI) GenBank. The recovery of DNA sequences from the herbal products was 60.4%, of which 81.97% were identified, and 18.03% showed no sequence through Basic Local Alignment Search Tool (BLAST) identification. Specific PCR combined with digestion using the restriction enzyme of Mbol allowed for identifying O. stamineus and E. longifolia. In contrast, Mspl allowed the identification of L. pumila by producing specific restriction patterns of plant samples. As a conclusion, a reliable approach for identifying and validating plant species in herbal products has been created using restriction enzymes. This simple and accurate PCR-RFLP approach efficiently identifies E. longifolia, L. pumila, and O. stamineus by analysing ITS2 sequences, assuring consumer health and safety. |
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