Sains Malaysiana 46(6)(2017): 909–915

http://dx.doi.org/10.17576/jsm-2017-4606-10

 

Kesan Penyimpanan Sampel Air Liur Terhadap Kualiti DNA Genom

(Storage Effect of Saliva Sample on Quality of Genomic DNA)

ROHAYA MEGAT ABDUL WAHAB¹, FARAH AMIRAH MOHD NASRI², INTAN ZARINA ZAINOL ABIDIN³, ZAIDAH ZAINAL ARIFFIN^4,5, MUHAMMAD DAIN YAZID⁶ & SHAHRUL HISHAM ZAINAL ARIFFIN^2*

 

¹Jabatan Ortodontik, Fakulti Pergigian, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Wilayah Persekutuan, Malaysia

 

²Pusat Pengajian Biosains dan Bioteknologi, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

 

³Pusat Sumber Penyelidikan, Cyberjaya University College of Medical Sciences, 63000 Cyberjaya, Selangor Darul Ehsan, Malaysia

 

⁴Jabatan Biologi, Fakulti Sains Gunaan, Universiti Teknologi MARA, 40450 Shah Alam, Selangor Darul Ehsan, Malaysia

 

⁵Atta-ur Rahman, Institut Kajian Ubat Semulajadi, Fakulti Farmasi, Universiti Teknologi MARA, 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia

 

⁶Pusat Kejuruteraan Tisu, Tingkat 12, Blok Klinikal, Pusat Perubatan Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, 56000 Cheras, Kuala Lumpur, Wilayah Persekutuan, Malaysia

 

Received: 3 July 2016/Accepted: 20 December 2016

 

ABSTRAK

Air liur berpotensi menjadi punca DNA yang mudah diambil bagi kajian klinikal kerana tidak invasif berbanding sampel darah. Kajian ini dijalankan untuk memencilkan dan menulenkan DNA genom daripada sampel air liur manusia serta mengkaji kesan penyimpanan terhadap kualiti DNA genom. Sampel air liur (n=5) disimpan dalam penimbal Tris-NaCl EDTA (TNE) pada suhu bilik (25°C) mengikut tempoh masa yang ditetapkan iaitu, segar (tanpa penyimpanan), 1,2,3 dan 4 bulan. Pemencilan dan penulenan DNA dilakukan menggunakan kaedah fenol-kloroform. Seterusnya, PCR telah dijalankan untuk mengetahui ketulenan DNA yang diekstrak menggunakan amplifikasi pada kawasan jujukan beta-globin dan mengenal pasti kehadiran bakteria melalui jujukan yang mengekod 16S rDNA. Keputusan menunjukkan fragmen DNA gen beta-globin manusia hanya berjaya diamplifikasi daripada sampel segar. Sampel air liur yang disimpan dalam penimbal TNE pada suhu bilik tidak mampu menstabilkan DNA genom manusia untuk jangka masa lama dan hanya berkesan untuk tempoh yang singkat iaitu, kurang daripada 1 bulan. Kesimpulannya, hanya sampel air liur segar sahaja yang berupaya memencil DNA genom.

 

Kata kunci: Ketulenan DNA; PCR; penimbal TNE; suhu bilik

 

ABSTRACT

Saliva is a potential source of DNA easily obtained for clinical studies because it is non-invasive compared to blood samples. This study was carried out to isolate and purify genomic DNA from human saliva sample and to study storage effect on the quality of genomic DNA. Saliva samples (n=5) were kept in Tris-NaCl EDTA (TNE) buffer at room temperature (25°C) according to fixed time period which are; fresh (without storage), 1,2,3 and 4 months. Isolation and purification of DNA was carried out using phenol-chloroform method. Next, PCR was conducted to determine the purity of extracted DNA by using amplification of beta-globin sequence region and identify bacterial existence using the sequence that codes for 16S rDNA. Only human beta-globin genomic DNA fragment was successfully amplified from fresh sample. Saliva sample kept in TNE buffer at room temperature was not able to stabilize human genomic DNA at long term and worked for short term storage which was less than a month. In conclusion, fresh saliva sample is needed to isolate genomic DNA.

 

Keywords: DNA purity; PCR; room temperature; TNE buffer

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*Corresponding author; email: shahroy8@gmail.com