Charunee Maharat, Uamanat Intaraphad, and Pojchanad Jantarasamee

pp. 993 - 1002

Abstract

The polymerase chain reaction (PCR) was used to determine pig DNA fragment in meat mixture
using specific oligonucleotide primer having 10-30 bp in size to amplify the DNA fragment. Firstly, the β-actin
F and the β-actin R primers were used to identify DNA extraction process in beef, pork and chicken. The 284
bp DNA fragment was obtained from beef and chicken meat and 248 bp was amplified from pork. Secondly,
pig F and pig R primers for pork were used as the PCR amplifier which resulted in 531 bp DNA fragment
from pork; however, it gave negative results for beef and chicken meat. Finally, the obtained nucleotide
sequence was compared with AF535163 in the GenBank database and homology was 98%. This work was
also submitted to GenBank and obtained an accession number of AY621117.
A comparison between the sensitivity of the commercial kit (QIAamp kit) and the RSB lysis buffer
for the DNA extraction process was carried out by mixing pork with chicken meat at ratios of 1:1, 1:5, 1:10, 1:50, 1:150 and 1:200. QIAamp kit gave the best results at the smallest ratio of 0.5% (1:200), while the RSB
lysis buffer gave good results at the ratio of 2% (1:50). This indicated that the QIAamp kit had a higher
sensitivity than the RSB lysis buffer. Lastly, a determination of pork DNA fragment from heated pork at
121ºC for 15 min. using the Pig F and Pig R primers for pork were done. The result was similar to that
obtained from the fresh pork at 531 bp DNA fragment.