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PCR-RFLP and Sequencing of trnS/trnfM Fragment of Enhalus acoriodes from Sanur Coastal Waters, Bali, Indonesia: A Preliminary Study Pharmawati, Made; Imaniar, Eka Fibayani
Journal of Tropical Life Science Vol 6, No 2 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.02.10

Abstract

This study aimed to evaluate the resolution of PCR-RFLP and sequencing of trnS/trnfM in detecting genetic diversity of seagrass Enhalus acoroides at Sanur Beach, Sindhu Beach and Semawang Beach. This research used six samples E. acoroides from each location. DNA extraction method followed Doyle and Doyle. PCR amplifications were done using primers P672/P673, P674/P675 and ITS4/ITS5 followed by digestion using restriction enzymes EcoRI, PstI, HindIII, BamHI, Rsa, Mva and HinfI. Sequencing analysis used PCR products of primers trnS/trnfM. Testing of DNA sequences of E. acoroides were conducted using BLAST (Basic Local Alignment Search Tool). DNA sequences were further analyzed using MEGA 5.2 software (Molecular Evolutionary Genetic Analysis) to evaluate variations of DNA. The sequence alignments were done using ClustalW software to determine the homology between the DNA sequences. The results showed that 18 samples of E. acoroides from Sanur, Sindhu and Semawang Beach have no polymorphism based on restriction enzyme analyses. Furthermore, sequencing of trnS/trnfM region of 18 samples E. acoroides showed that the sequences were identical.   
PCR-RFLP and Sequencing of trnS/trnfM Fragment of Enhalus acoriodes from Sanur Coastal Waters, Bali, Indonesia: A Preliminary Study Made Pharmawati; Eka Fibayani Imaniar
Journal of Tropical Life Science Vol. 6 No. 2 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.02.10

Abstract

This study aimed to evaluate the resolution of PCR-RFLP and sequencing of trnS/trnfM in detecting genetic diversity of seagrass Enhalus acoroides at Sanur Beach, Sindhu Beach and Semawang Beach. This research used six samples E. acoroides from each location. DNA extraction method followed Doyle and Doyle. PCR amplifications were done using primers P672/P673, P674/P675 and ITS4/ITS5 followed by digestion using restriction enzymes EcoRI, PstI, HindIII, BamHI, Rsa, Mva and HinfI. Sequencing analysis used PCR products of primers trnS/trnfM. Testing of DNA sequences of E. acoroides were conducted using BLAST (Basic Local Alignment Search Tool). DNA sequences were further analyzed using MEGA 5.2 software (Molecular Evolutionary Genetic Analysis) to evaluate variations of DNA. The sequence alignments were done using ClustalW software to determine the homology between the DNA sequences. The results showed that 18 samples of E. acoroides from Sanur, Sindhu and Semawang Beach have no polymorphism based on restriction enzyme analyses. Furthermore, sequencing of trnS/trnfM region of 18 samples E. acoroides showed that the sequences were identical.   
Co-Authors A. A. Putri Ardyanti A.A.S. A. Sukmaningsih Agriani Dewinta Anak Agung Ayu Putri Ardyanti Anak Agung Gde Raka Dalem Andi Bahtiar Batti Andrianus Sembiring, Andrianus Aziz, Alifya Ibnu Bayu Adjie Bayu Adjie Bayu Adjie Bram Setyadji Deny Suhermawan Yusup Dian Catur Prayantini Dwi Nur Rikhma Sari Eka Fibayani Imaniar Eka Fibayani Imaniar Ema Hendriyani Fransiska Nitti Fransiskus Jimmy Roga Gde Evayanti, Luh GUIJUN YAN Gusti Ayu Putri Arnila I Gde Suryadi I Gede Putu Ardhana I Gusti Agung Eka Saraswati I Gusti Ayu Agung Pradnya Paramitha I Gusti Ngurah Kade Mahardika I Ketut Catur Wiguna I Ketut Junitha I Ketut Suada I Komang Adi Widyastama I MADE AGUS WIARTANA I Made Anom Sutrisna Wijaya I Made Murna I Putu Agus Tirta Cahyana I Putu Satwika I Wayan Arnata I Wayan Rosiana I. Setyawati I.G.A.S. Wahyuni I.M.A.S. Wijaya IAN JAMES MACFARLANE IDA AYU ASTARINI Ida Ayu Ratih Purnama Dewi Imaniar, Eka Fibayani Indah Tria Hoky Inna Narayani IRIANI SETYAWATI Joko Raharjo, Sentot Junita Hardini Ketut Queena Fredlina Luh Putu Wrasiati Made Ria Defiani Maharani, Rai Janaki Maliza Kurnia Maya Agustina Maya Agustina Mery Wintari Ngurah Intan Wiratmini Ni Kadek Dewi Rustini NI KADEK YUNITA SARI Ni LUH ARPIWI Ni Luh Ayu Jami Wistiani, Ni Luh Ayu Jami Ni Luh Putu Agustini Ni Luh Watiniasih Ni Luh Wayan Yulia Mirayanti NI MADE DIAN PRATIWI Ni Made Gari Ni Made Sastriyani Wiendra Ni Made Sastriyani Wiendra Ni Made Suartin Ni Nyoman Wirasiti Ni Putu Adriani Astiti Ni Putu Eka Krisnayanti Ni Wayan Ayu Wiartini Ni Wayan Karolina Ni Wayan Sudatri PATRICK FINNEGAN Putera, I Kadek Aridena Putu Angga Wiradana Putu Apriliani Putu Yunita Putri Aryani Retno Kawuri Rifky Firmansyah Anwar Satwika, I Putu Septarini Dian Anitasari Septarini Dian Anitasari Siddique, Kadambot.H.M. Tassanai Jaruwatanaphan Tassanai Jaruwatanaphan Tassanai Jaruwatanaphan, Tassanai Tiwi Wati Uslan UUL SHOVI NURKAMILA Vanesa Martida Vanesa Martida Viryanando Evan Rahardja WAHYU WIDOWATI Wawan Sujarwo WENNI SETYO LESTARI Wenni Setyo Lestari Wenni Setyo Lestari, Wenni Setyo Wiguna, I Ketut Catur Wijaya, Putu Austin Widyasari Wildan Mujahidul Basyar Yasuyuki Watano Yasuyuki Watano Yasuyuki Watano, Yasuyuki Yenni Ciawi Zaetuna, Hanifa Laila