Malaysian Applied Biology Journal

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46_04_08

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Malays. Appl. Biol. (2017) 46(4): 63–71

OPTIMISATION OF POLYMERASE CHAIN REACTION

CONDITIONS TO AMPLIFY D-LOOP REGION IN THE

MALAYSIAN MOUSEDEER GENOMIC DNA

 

MOHAMAD-AZAM AKMAL ABU BAKAR1, JEFFRINE JAPNING ROVIE-RYAN2,

AHMAD AMPENG3, SALMAH YAAKOP1, SHUKOR MD NOR1

and BADRUL MUNIR MD-ZAIN1*


1School of Environmental and Natural Resource Sciences, Faculty of Science and Technology,

Universiti Kebangsaan Malaysia, 43600, Selangor, Malaysia

2Department of Wildlife and National Parks (PERHILITAN), Peninsular Malaysia,

Kuala Lumpur, Malaysia

3Sarawak Forest Department, Petra Jaya, Kuching Sarawak

 

*Email: This e-mail address is being protected from spambots. You need JavaScript enabled to view it , This e-mail address is being protected from spambots. You need JavaScript enabled to view it

 

Accepted 27 November 2017, Published online 31 December 2017

 

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 ?M 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.

Key words: Polymerase chain reaction, D-loop, annealing temperature, mousedeer, optimisation

 

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