DNA isolation and optimization of ISSR-PCR reaction system in Oryza sativa L.

Azhar Mohamad, Arshad Naji Alhasnawi, Ahsan A. Kadhimi, Anizan Isahak, Wan Mohtar Wan Yusoff, C. M.Z.Che Radziah

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Inter simple sequence repeats (ISSRs) have been utilized widely for molecular markers in analyzing the genetic diversity and phylogenetic and regions in the genome flanked by microsatellite sequences. PCR amplification of these regions using a single primer yields multiple amplification products that can be used as a dominant multilocus marker system for the study of genetic variation in various organisms. For this study provides, DNA isolation, adjusting in six factors (Buffer, MgCl2, dNTPs, ISSR primers, Template DNA and Taq polymerase) at six levels, and optimization of PCR temperature for the ISSR reaction was 60-45 °C, primers screening on indica rice (Oryza sativa). In this research, simple method of DNA isolation by using seedling. The objective of the present investigation was to assess the optimizations and quantification. Has been shown that stalk enhanced the maximum value of genomic. The results show that 100 ISSR primers were examined as well as, 56 ISSR primers was productively amplified. Optimum components for PCR reactions were 5.0 μl of 5X PCR Buffer, 1.5 μl of 25mM MgCl2, 1 μl of 10 mM dNTP, 1 μl of 10 Mm ISSR primers, 2 μl Template DNA, and 0.1 μl of 5 units/ml Taq polymerase. Based on this study, has brought out some information on the relationship between these ISSR primers will be applied further for molecular profiling as well as response evaluation in rice varieties.

Original languageEnglish
Pages (from-to)2264-2272
Number of pages9
JournalInternational Journal on Advanced Science, Engineering and Information Technology
Volume7
Issue number6
Publication statusPublished - 1 Jan 2017

Fingerprint

Microsatellite Repeats
Oryza sativa
DNA
microsatellite repeats
Polymerase Chain Reaction
Amplification
Taq Polymerase
Magnesium Chloride
Buffers
Screening
buffers
Genes
rice
genetic variation
Oryza
DNA primers
Seedlings
Genome
screening
genomics

Keywords

  • DNA isolation
  • ISSR screening
  • Optimal PCR condition
  • Plant genetics

ASJC Scopus subject areas

  • Computer Science(all)
  • Agricultural and Biological Sciences(all)
  • Engineering(all)

Cite this

Mohamad, A., Alhasnawi, A. N., Kadhimi, A. A., Isahak, A., Wan Yusoff, W. M., & Radziah, C. M. Z. C. (2017). DNA isolation and optimization of ISSR-PCR reaction system in Oryza sativa L. International Journal on Advanced Science, Engineering and Information Technology, 7(6), 2264-2272.

DNA isolation and optimization of ISSR-PCR reaction system in Oryza sativa L. / Mohamad, Azhar; Alhasnawi, Arshad Naji; Kadhimi, Ahsan A.; Isahak, Anizan; Wan Yusoff, Wan Mohtar; Radziah, C. M.Z.Che.

In: International Journal on Advanced Science, Engineering and Information Technology, Vol. 7, No. 6, 01.01.2017, p. 2264-2272.

Research output: Contribution to journalArticle

Mohamad, A, Alhasnawi, AN, Kadhimi, AA, Isahak, A, Wan Yusoff, WM & Radziah, CMZC 2017, 'DNA isolation and optimization of ISSR-PCR reaction system in Oryza sativa L.', International Journal on Advanced Science, Engineering and Information Technology, vol. 7, no. 6, pp. 2264-2272.
Mohamad, Azhar ; Alhasnawi, Arshad Naji ; Kadhimi, Ahsan A. ; Isahak, Anizan ; Wan Yusoff, Wan Mohtar ; Radziah, C. M.Z.Che. / DNA isolation and optimization of ISSR-PCR reaction system in Oryza sativa L. In: International Journal on Advanced Science, Engineering and Information Technology. 2017 ; Vol. 7, No. 6. pp. 2264-2272.
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