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EKSTRAKSI DAN AMPLIFIKASI DNA RARU Cotylelobium Melanoxylon Pierre DAN Cotylelobium Lanceolatum Craib

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EKSTRAKSI DAN AMPLIFIKASI DNA RARU Cotylelobium Melanoxylon Pierre DAN Cotylelobium Lanceolatum Craib EKSTRAKSI DAN AMPLIFIKASI DNA RARU Cotylelobium melanoxylon Pierre DAN Cotylelobium lanceolatum Craib SKRIPSI PUTRI GEA 151201059 DEPARTEMEN BUDIDAYA HUTAN FAKULTAS KEHUTANAN UNIVERSITAS SUMATERA UTARA MEDAN 2020 UNIVERSITAS SUMATERA UTARA EKSTRAKSI DAN AMPLIFIKASI DNA RARU Cotylelobium melanoxylon Pierre DAN Cotylelobium lanceolatum Craib SKRIPSI Oleh : PUTRI GEA 151201059 Skripsi sebagai salah satu syarat untuk memperoleh gelar sarjana di Fakultas Kehutanan Universitas Sumatera Utara DEPARTEMEN BUDIDAYA HUTAN FAKULTAS KEHUTANAN UNIVERSITAS SUMATERA UTARA MEDAN 2020 UNIVERSITAS SUMATERA UTARA i UNIVERSITAS SUMATERA UTARA ii UNIVERSITAS SUMATERA UTARA ABSTRAK PUTRI GEA : Ekstraksi dan Amplifikasi DNA Raru Cotylelobium melanoxylon Pierre dan Cotylelobium lanceolatum Craib. Di bawah bimbingan ARIDA SUSILOWATI dan HENTI HENDALASTUTI RACHMAT. Raru (Cotylelobium melanoxylon Pierre) dan (Cotylelobium lanceolatum Craib) merupakan pohon yang tumbuh pada daerah beriklim tropis dan tumbuh tersebar di Pulau Sumatera dan Kalimantan. Pemanenan pohon yang tidak terkontrol telah menyebabkan penurunan populasi alami yang cepat menyebabkan potensi penurunan keanekaragaman genetik. Informasi terkait dengan genetika raru di Sumatera Utara masih sangat terbatas, termasuk metode ekstraksi DNA yang sesuai. Oleh karena itu, penelitian ini dilakukan untuk mendapatkan informasi tentang ekstraksi DNA menggunakan metode CTAB dan penanda referensi untuk studi genetik populasi. Ekstraksi DNA dilakukan melalui metode CTAB (Cetyl Trimetyl Ammonium Bromide). DNA hasil ekstraksi selanjutnya diamplifikasi dengan PCR menggunakan tiga penanda standar barkoding yaitu ITS, trnH-psbA dan trnL-trnF. Hasil penelitian menunjukkan bahwa metode CTAB menghasilkan kualitas DNA yang kurang baik dan pengenceran 10% memberikan hasil amplifikasi produk PCR terbaik. Primer trnL-trnF mampu mengamplifikasi DNA raru dengan suhu annealing 50ºC dan trnH- psbA 54ºC. Kata Kunci : amplifikasi, DNA, ekstraksi, genetik, raru iii UNIVERSITAS SUMATERA UTARA ABSTRACT PUTRI GEA: DNA Extraction and Amplification of Raru Cotylelobium melanoxylon Pierre and Cotylelobium lanceolatum Craib Supervised by ARIDA SUSILOWATI and HENTI HENDALASTUTI RACHMAT. Cotylelobium melanoxylon Pierre and Cotylelobium lanceolatum Craib or locally known as raru,is tropical tree species and commonly distributed on the Sumatera and Kalimantan Island. Uncontrolled harvesting of wild-grown trees has led to rapid decrease of their natural populations and causing potential decrease in genetic diversity. The information on genetic aspect on raru in North Sumatera still limited, including it extraction methods.Therefore, this research was conducted to get information about optimal DNA isolation using CTAB methods and reference marker for further used in genetic study. DNA extraction was conducted through CTAB (Cetyl Trimetyl Ammonium Bromide) method. The isolation product then amplified using three barcoding standard marker those were ITS, trnH-psbA and trnL-trnF. The result showed that CTAB method was not able to yield good quality DNA and 10% dilution produced the best amplification of PCR product. The trnL-trnF primer was able to amplified DNA of raru with 50ºC annealing temperature and trnH-psbA with 54ºC annealing temperature. Keywords:,amplification, DNA,extraction, genetic, raru. iv UNIVERSITAS SUMATERA UTARA RIWAYAT HIDUP Penulis lahir di Kota Gunungsitoli pada tanggal 07 Januari 1998. Penulis merupakan anak tunggal dari pasangan Bapak Ridwan Gea dan Ibu Bakhtiar Gea. Pada Tahun 2009 penulis lulus dari SDN 070981 Fodo. Penulis kemudian melanjutkan studi ke SMP Negeri 1 Gunungsitoli dan lulus pada tahun 2012. Penulis lulus pada tahun 2015 dari MA Negeri Gunungsitoli. Pada tahun 2015, penulis diterima di Universitas Sumatera Utara (USU) melalui jalur Seleksi Nasional Masuk Perguruan Tinggi Negeri (SNMPTN) sebagai mahasiswa di Program Studi Kehutanan Fakultas Kehutanan, Universitas Sumatera Utara. Selain mengikuti perkuliahan, penulis menerima beasiswa BIDIKMISI sejak tahun 2015. Penulis juga aktif mengikuti kegiatan organisasi sebagai anggota divisi DANUS (Dana dan Usaha) dalam organisasi BKM Baytul Asyjar dan GORGA (Gerakan Observasi Rimbawan Giat Alam) dari tahun 2017-2018 di Fakultas Kehutanan, Universitas Sumatera Utara serta mengikuti kegiatan organisasi PMIN-Medan dan menjabat sebagai Sekretaris Bidang Pembinaan dan Pengembangan Anggota dari tahun 2017-2018. Penulis melakukan Praktik Pengenalan Ekosistem Hutan (P2EH) di Kawasan Hutan Dengan Tujuan Khusus (KHDTK) Pondok Buluh pada tahun 2016. Penulis melaksanakan Praktik Kerja Lapangan (PKL) di Pusat Penelitian dan Pengembangan Hutan Bagian Kerjasama Komatsu-FORDA Bogor pada tanggal 23 Juli sampai 23 Agustus 2018. v UNIVERSITAS SUMATERA UTARA KATA PENGANTAR Puji syukur penulis ucapkan kehadirat Allah SWT atas rahmat dan karunia-Nya penulis dapat menyelesaikan skripsi ini. Skripsi ini berjudul “Ekstraksi dan Amplifikasi DNA RARU Cotylelobium melanoxylon Pierre dan Cotylelobium lanceolatum Craib”. Penulisan skripsi ini merupakan syarat dalam mencapai gelar sarjana di Fakultas Kehutanan, Universitas Sumatera Utara. Penulis banyak menerima bimbingan, motivasi, saran dan juga doa dari berbagai pihak selama penulisan skripsi ini. Teristimewa dari kedua orang tua yang sangat penulis sayangi yaitu Ayahanda Ridwan Gea dan Ibunda Bakhtiar Gea yang tidak pernah berhenti memberikan kasih sayang, doa, dukungan, juga nasihat yang tulus sampai sekarang ini. Pada kesempatan ini penulis mengucapkan terima kasih kepada : 1. Ibu Dr. Arida Susilowati, S.Hut., M.Si dan Ibu Henti Hendalastuti Rachmat, S.Hut., M.Si., Ph.D selaku komisi pembimbing yang telah membimbing dan mengarahkan penulis serta memberikan berbagai masukan berharga kepada penulis dalam penyelesaian skripsi ini. 2. Ibu Dr. Evalina Herawati, S.Hut., M.Si sebagai Dosen Penguji I, Ibu Ridahati Rambey, S.Hut,. M.Si, sebagai Dosen Penguji II dan Bapak Dr. Muhdi S.Hut., M.Si sebagai Dosen Penguji III. 3. Ibu Siti Latifah, S.Hut., M.Si.,Ph.D selaku Dekan Fakultas Kehutanan Universitas Sumatera Utara. 4. Bapak Mohammad Basyuni, S.Hut., M.Si., Ph.D sebagai Ketua Departemen Budidaya Hutan dan Ibu Dr. Deni Elfiati, SP., MP selaku Sekretaris Departemen Budidaya Hutan Fakultas Kehutanan Universitas Sumatera Utara. 5. Bapak Ahmad Baiquni Rangkuti, S.Hut., M.Si yang bersedia memberikan dukungan materi dan moral untuk pelaksanaan dan penyusun hasil penelitian ini. 6. Laboratorium Genetik dan Molekuler Kehutanan, Dapartemen Silvikultur, Fakultas Kehutanan, Institut Pertanian Bogor yang telah memberi ijin sebagai lokasi penelitian dan atas semua bantuan yang telah diberikan kepada peneliti. 7. Seluruh sahabat penulis yaitu Tim Penelitian, Gadis Sampul, Tim PKL Litbang Bogor, HUT B 2015, BDH 2015 dan seluruh teman teman dan adik-adik junior yang telah membantu dan memberi semangat dalam proses penelitian. Penelitian ini terlaksanakan atas bantuan hibah penelitian dosen dari dana hibah Pusat Penelitian dan Pengembangan Hutan Bagian Kerjasama Komatsu-FORDA Bogor penulis berharap semoga skripsi ini memberikan manfaat ke berbagai pihak. Akhir kata penulis mengucapkan terima kasih. Medan, Januari 2020 Putri Gea vi UNIVERSITAS SUMATERA UTARA DAFTAR ISI Halaman LEMBAR PENGESAHAN ............................................................................. i PERNYATAAN ORIGINALITAS ................................................................. ii ABSTRAK ....................................................................................................... iii ABSTRACT ....................................................................................................... iv RIWAYAT HIDUP .......................................................................................... v KATA PENGANTAR ..................................................................................... vi DAFTAR ISI .................................................................................................... vii DAFTAR TABEL ............................................................................................ ix DAFTAR GAMBAR ....................................................................................... x DAFTAR LAMPIRAN .................................................................................... xi PENDAHULUAN Latar Belakang................................................................. ................................ 1 Tujuan Penelitian....................................................... ...................................... 3 Kegunaan Penelitian......................................................................................... 4 TINJAUAN PUSTAKA Taksonomi Pohon Cotylelobium ...................................................................... 4 Desripsi Umum Pohon Cotylelobium .............................................................. 4 Sebaran Pohon Cotylelobium ........................................................................... 6 Manfaat Pohon Cotylelobium ........................................................................... 6 Permasalahan Pohon Cotylelobium .................................................................. 6 Penanda Molekuler........................................................................................... 6 Ekstraksi DNA ................................................................................................. 7 Metde Cetyl Trimethyl Ammonium Bromide (CTAB) ..................................... 8 Elektroforesis ..................................................................................................
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