Optimisation of polymerase chain reaction conditions to amplify D-loop region in the Malaysian mousedeer genomic DNA

Mohamad Azam Akmal Abu Bakar, Jeffrine Japning Rovie-Ryan, Ahmad Ampeng, Salmah Yaakop, Shukor Md. Nor, Badrul Munir Md. Zain

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Polymerase chain reaction (PCR) has been utilized in molecular studies to amplify DNA fragments even from small amounts of genetic material. A good PCR product is based on good amplification of DNA segments in condition optimisation. We conducted a molecular study to optimize the annealing temperature and concentration of primers to find the optimal condition of PCR for Malaysian mousedeer species. DNA samples from six Tragulus kanchil and Tragulus napu species of Malaysian mousedeer were extracted for PCR amplification. Several ranges of annealing temperatures and concentrations were tested using the increment and decrement method. Results indicated that the optimal annealing temperatures for T. kanchil and T. napu were 56.0ºC and 47.2ºC, respectively. For primer concentration, 10 to 20 μM is best to produce good PCR amplification. All six samples for the D-loop region had good DNA chromatograms and scores between 90% and 100% in BLAST. Based on the high scores, all mousedeer samples were considered to be highly purified DNA.

Original languageEnglish
Pages (from-to)63-71
Number of pages9
JournalMalaysian Applied Biology
Volume46
Issue number4
Publication statusPublished - 1 Dec 2017

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polymerase chain reaction
annealing
genomics
Polymerase Chain Reaction
DNA
Temperature
temperature
sampling
Genes
methodology

Keywords

  • Annealing temperature
  • D-loop
  • Mousedeer
  • Optimisation
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Optimisation of polymerase chain reaction conditions to amplify D-loop region in the Malaysian mousedeer genomic DNA. / Bakar, Mohamad Azam Akmal Abu; Rovie-Ryan, Jeffrine Japning; Ampeng, Ahmad; Yaakop, Salmah; Md. Nor, Shukor; Md. Zain, Badrul Munir.

In: Malaysian Applied Biology, Vol. 46, No. 4, 01.12.2017, p. 63-71.

Research output: Contribution to journalArticle

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AU - Yaakop, Salmah

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