RAPD Primers Selection for Genetic Variation Analysis of Banana Plant (Musa spp.)
DOI:
https://doi.org/10.25077/jbioua.9.2.47-53.2021Keywords:
Genetic, DNAAbstract
Identification of genetic diversity using molecular markers is important for basic information for plant conservation. Banana is a fruit bearing plants that important for food sources in human life. This study aimed to determine the optimum conditions of  PCR-RAPD reaction and RAPD primers that suitable to amplify DNA fragmens. DNA isolation was done using modified of CTAB and chloroform isoamil alcohol. The samples used was young leaves of nine banana cultivar plants. Optimation was done using variety of DNA and MgCl2 concentration. Eight primers produced by Operon Primer Technology were tested. The DNA genomic concentration obtained was in the range of 23,3 ng/µl – 70 ng/µl. The optimum conditions of PCR-RAPD of banana plants that produce clear band were 50 ng/µl DNA template, 3 mM MgCl2 with the number of thermal cycles was 40 x. There were six RAPD primers that successfully amplified DNA : OPA 02, OPA 04, OPB 12, OPD 20, OPH 01, and OPH 03. The primer OPA-04 had the lowest resolving power value (4,4) , while  OPH 01 had the highest (11,3) resolving power.
References
Ali, B.A., Huang, T.H., Salem, H.H. and Xie, Q.D. 2006. Influence of Thermal Cycler Day to Day Reproducibility of Random Amplified Polymorphic DNA Fingerprints. Biotechnology 5 : 324-329
Choudhary, R., Keshavachandran, R., Menon, R., Khalekar, G., Singh, N., and Maruthiyottu, D. 2014. Molecular Variability of Plantain Ecotypes from the Genus Musa (Musaceae). Turkish Journal of Botany 38 : 827-834.
Doyle, J.J., and Doyle, J.L. 1990. Isolation of Plant DNA from Fresh Tissue. Focus 12 : 13-15.
Eurofinsgenomic. 2014. RAPD 10mer KITS. https://eurofinsgenomics.eu/media/1610370/rapd_10mer_kits_sequences.pdf
Innis M. A., Gelfand D. H., Sninsky J.J. and White T.J. 1990. PCR Protocols, A Guide to Methods and Applications. New York : Academic Press Incoporated.
Kasijadi, F. 2006. Penerapan Agribisnis Berbasis Pisang Spesifik Lokasi Pisang Mas dan Agung. Jawa Timur : Balai Pengkajian Teknologi Pertanian.
Lorenz, T. C. 2012. Polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies. Journal of visualized experiments : JoVE (63), e3998. https://doi.org/10.3791/3998
Martida, V., dan Pharmawati, M. 2016. Pemilihan Primer RAPD (Random Amplified Polymorphic DNA) pada PCR (Polymerase Chain Reaction) Tanaman Kamboja (Plumeria sp.). Sombiosis 4(1):16-18
Nadeem, M.A., Nawaz, M.A., Shahid, M.Q., , Doğan,Y., Comertpay G., Yıldız M., Hatipoğlu R., Ahmad, F., Alsaleh, A., Labhane N., Özkan H., Chung G., and Baloch F.S. 2018. DNA molecular markers in plant breeding: current status and recent advancements in genomic selection and genome editing. Biotechnology and Biotechnological Equipment 32(2):261–285.
Pharmawati, M. 2009. Optimalisasi Ekstraksi DNA dan PCR-RAPD pada Grevillea spp. Jurnal Biologi 13 (1) : 12-16.
Prayitno, E. dan Nuryandani E. 2011. Optimalisasi Ekstraksi DNA Jarak Pagar (Jatropha curcas) Melalui Pemilihan Daun yang Sesuai. Bioteknologi 8 (1) : 24-31.
Prevost, A., Wilkinson, M.J. 1999. A New System of Comparing PCR Primers Applied to ISSR Fingerprinting of Potato Cultivars. Theoretical and Applied Genetics 98 : 107–112.
Randriani, E., Tresniawati, C., dan Syafaruddin. 2012. Pemanfaatan Teknik Random Amplified Polymorphic DNA (RAPD) Unruk Pengelompokan Secara Genetik Plasma Nutfah Jambu Mete (Anacardium occidentale L.). Buletin RISTRI 3 (1) : 1-6.
Sambrook , J. and Russell, D. W. 2001. Molecular Cloning: A Laboratory Manual. Third Edition. New York : Cold Spring Harbor Laboratory Press.
Saraswathi, U., and Mullainathan, L. 2020. Extraction of high-quality genomic DNA and identification of different DNA barcoding markers for chickpea (Cicer arietinum L.). Buletin Agroteknologi 1(1):7-11
Usha, K., Moahnty, S.K., Roy, P.S, Behera, L. And Chand, P.K. 2015. Genetic Diversity among Banana Cultivars from Odisha using RAPD Markers. Science Research Article 5 (2) : 118-124.
Uslan dan Pharmawati, M. 2015. Optimasi Konsentrasi DNA dan MgCl2 pada Reaksi Polymerase Chain Reaction-Random Amplified Polymorphic DNA untuk Analisis Keragaman Genetik Tanaman Faloak (Sterculia quadrifida R.Br). Jurnal Bioslogos. 5 (1) : 26-34.
Williams, J. G., Kubelik, A.R., Livak, K. J., Rafalski, J.A. and Tingey, S.J. 1990. DNA Polymorphisms Amplified by Arbitrary Promers are Useful as Genetic Markers. Nucleid Acids Research 18 : 6531-6535.
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