STANDARDISASI BAHAN HERBAL ANTIDIABETES DAUN Gyrinops versteegii SECARA MOLEKULAR MENGGUNAKAN SEKUEN rpoC1

I Gusti Agung Ayu Hari Triandini, I Gde Adi Suryawan Wangiyana

Abstract


Penelitian ini bertujuan untuk melakukan analisis molekular bahan herbal antidiabetes daun gaharu (Gyrinops versteegii) menggunakan sekuen rpoC1. Daun G. versteegii diambil dari perkebunan gaharu di wilayah Lingsar. Daun diolah menjadi bahan herbal dengan menggunakan 4 SOP: pencucian, pengeringan, pencacahan, dan oksidasi. Isolasi DNA dilakukan dengan menggunakan Blood – Animal DNA Preparation Kit sesuai protokol dari produsen. DNA genom diukur absorbansinya pada panjang gelombang 230 nm, 260 nm, dan 280 nm. Amplifikasi PCR dilakukan dengan menggunakan primer universal rpoC1 dengan program initial denaturation 95oC selama 3 menit diikuti 40 siklus denaturasi 95oC selama 15 detik, annealing 37oC selama 1 menit, extension 72oC selama 2 menit, dan final extension 72oC selama 5 menit. MegaBLAST dari laman NCBI digunakan dalam pencarian sekuen referensi. Pohon filogenetik direkonstruksi dengan menggunakan 2 algoritme: Neighbor Joining dan Maximum Parsimony. Hasil penelusuran MegaBLAST menunjukkan bahwa anggota genus Aquilaria mendominasi database referensi sekuen dengan score tertinggi. Pohon filogenetik Neighbor Joining dan Maximum Parsimony yang direkonstruksi dari referensi sekuen rpoC1 memiliki topology yang sedikit berbeda. Meskipun demikian, pada kedua pohon filogenetik tersebut, G. versteegii selalu tergabung dalam satu clade dengan spesies Aquilaria. Dapat disimpulkan bahwa Analisis molekular terhadap bahan baku antidiabtes dari daun G. versteegii menggunakan sekuen rpoC1 menghasilkan data bahwa gaharu G. versteegii secara molekular berhubungan dekat dengan spesies gaharu dari genus Aquilaria.

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DOI: https://doi.org/10.33394/jss.v5i2.7063

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