Sains Malaysiana 48(3)(2019): 555–559

http://dx.doi.org/10.17576/jsm-2019-4803-07

 

Successful and Efficient DNA Extraction from Processed Pangolin Scales

(Kejayaan dan Kecekapan Pengekstrakan DNA daripada Sisik Tenggiling Terproses)

 

XUAN CAI¹, HAO-TING YE¹, EN-WEI TIAN¹, FANG LI¹, CHAN LI¹, YAO-JUN YANG², CHUN-SHENG LIU² & ZHI CHAO^1*

 

¹School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China

 

²School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China

 

Diserahkan: 9 Mac 2018/Diterima: 14 November 2018

 

ABSTRACT

A successful method for total DNA extraction from crude and processed pangolin scales was established. After pretreatment in the soaking solution for cleansing, the scales were prepared into fine powders and treated with PBS buffer containing 0.1% collagenase and 0.1% trypsin for 24 h, followed by digestion with proteinase K at 55°C for 120 h. Phenol-chloroform extraction was used to obtain the total DNA. PCR amplification for mitochondrial cytochrome b (cytb) gene was successful using the extracted DNA as the template, and sequencing of the amplified fragments confirmed Manis origin of the scale samples. With an efficiency up to 100%, this method is expected to provide a powerful tool in molecular identification of processed as well as crude pangolin scales.

 

Keywords: DNA extraction; Manis; processed pangolin scales

 

ABSTRAK

Kaedah berkesan untuk pengekstrakan DNA total daripada sisik tenggiling mentah dan terproses telah berjaya dihasilkan. Selepas pra-rawatan dengan cara merendam di dalam larutan untuk pembersihan, sisik disediakan dalam bentuk serbuk halus dan dirawat dengan penimbal PBS yang mengandungi 0.1% kolagenase dan tripsin 0.1% selama 24 jam diikuti penghadaman oleh proteinase K pada 55°C selama 120 jam. Pengekstrakan fenol-kloroform telah digunakan untuk mendapatkan DNA total. Amplifikasi PCR untuk gen mitokondria sitokrom b (cytb) telah berjaya dengan menggunakan ekstrak DNA sebagai templat dan penjujukan serpihan teramplifikasi membuktikan sampel sisik berasal daripada Manis. Dengan kadar kecekapan sehingga 100%, kaedah ini dijangka menjadi suatu alat berkesan untuk mengenal pasti molekul sisik terproses serta mentah tenggiling.

 

Kata kunci: Manis; pengekstrakan DNA; sisik tenggiling terproses

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*Pengarang untuk surat-menyurat; email: chaozhi@smu.edu.cn