AMPLIFIKASI PCR DOMAIN D1/D2 28S rDNA MENGGUNAKAN PRIMER ITS1 DAN ITS4 SAMPEL DNA DARI Candida tropicalis YANG DIISOLASI DENGAN METODE PENDINGINAN

Authors

  • Hermansyah Hermansyah Jurusan Kimia MIPA Universitas Sriwijaya, Jalan Raya Palembang Prabumulih KM 32 Indralaya, Ogan Ilir, Sumatera Selatan
  • Novian Sutami Jurusan Kimia MIPA Universitas Sriwijaya, Jalan Raya Palembang Prabumulih KM 32 Indralaya, Ogan Ilir, Sumatera Selatan
  • Miksusanti Miksusanti Jurusan Kimia MIPA Universitas Sriwijaya, Jalan Raya Palembang Prabumulih KM 32 Indralaya, Ogan Ilir, Sumatera Selatan

DOI:

https://doi.org/10.26418/indonesian.v1i1.26037

Abstract

The purpose of this research was to isolated DNA from the yeast C. tropicalis with freeze thawing method -200 C conducted on 3 colonies of C. tropicalis.   Each colony     threated variations of cooling, 3x15 minutes, 3x25 minutes and 3x35 minutes, to break the cell walls.   Subsequently all the samples amplified with 3 variations of PCR cycles, 15 cycles, 25 cycles and 35 cycles, after all of the samples isolated by freeze thawing method -200 C. Its was known that sample A15 has the smallest concentration of DNA yeast C. tropicalis, ie 50 µg/mL, while sample C35 had the largest concentration of DNA yeast C. tropicalis, ie 225 µg/mL. The result of the research indicated that the best condition can be reached in 3x35 minutes. On 35th cycle has clearer C. tropicalis DNA bands than the 25th and 15th PCR cycle. C. tropicalis DNA bands at 35th cycles there were 7 DNA bands were detected and bright bands on a long 35 minutes cooling. In the 25th and the 15th cycle, there was no DNA bands were detected in all samples. Based on the results obtained, the amplification process must be carried out at least 35 times cycles so that the C. tropicalis DNA bands can be detected.

References

Jawetz. (2008). Mikrobiologi Kedokteran. Jakarta : EGC.

James, S.A and Stratford M. (2003). Spoilage yeast with emphasis on the genus. Woodhed publishing limited Cambridge. 171-191.

Fujita, S.I., Senda, Y., Nakaguchi, S., and Hashimoto, T. (2011). Multiplex PCR Using Internal Transcribed Spacer 1 and 2 Regions for Rapid Detection and Identi cation of Yeast Strains, Journal of Clinical Microbiology, 39, 10: 3617-3622.

Ciardo, G., Luttgen G. And Siminiceanu G. (2001). Strategy for symbolic state space generation. Construction and analysis of system. Italy : Springer verlag.

Kurtzman, C. P. and Robnett, C. J. (1998). Identiï¬cation and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek, 73 : 331–371. Microbiol Biotechnol. 43 : 242.

Retno A. (2008). Metode isolasi DNA. Jakarta : Universitas Indonesia.

Samaranayake, L.P. (2006) Essential microbiology for dentistry. USA : Elsevier limited.

Hermansyah., Novia., Sugiyama, M. and Harashima S. (2015). Candida tropicalis isolated from Tuak, a North Sumatera Indonesian Traditional Beverage, for Bioethanol Production.

Barnett, J.A., Payne R.W., and Yarrow D. (2000). Yeast. Characteristic and identification. Cambridge University Press, 9 : 11-39.

Rapley R. and Heptinstall J. (1998). UV spectrophotometric analysis of ribonucleic acids. Isolation and characterization protocols. Totowa : Humana Press Inc.

Ratnayani, K., Yowani, S.C. and Syane, L. (2009). Amplifikasi framen 0,4 kb daerah D-Loop DNA mitokondria lima individu suku Bali tanpa hubungan kekerabatan dengan metode PCR. Jurnal Kimia, 3 : 14-20.

Ethica, S.M., Nataningtyas, D.R., Lestari, P., Istini, Semiarti, E., Widada, J., and Raharjo, T.J. (2013). Comparative evaluation of conventional versus rapid methods for amplifiable genomic DNA isolation of cultured Azospirillium sp. JG. 3. Indo. J. Chem, 13 : 248-253.

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Published

2018-07-23

Issue

Vol. 1 No. 1 (2018)

Section

Articles

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Copyright (c) 2018 Hermansyah Hermansyah, Novian Sutami, Miksusanti Miksusanti

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