PIG SPECIES IDENTIFICATION IN

MEATBALLS USING POLYMERASE

CHAIN REACTION RESTRICTION
FRAGMENT LENGTH POLYMORPHISM

FOR HALAL AUTHENTICATION

Yuny Erwanto1, Mohammad Zainal Abidin1,
Abdul Rohman2 and Sismindari3

1Division of Animal Products Technology, Faculty of Animal Science, Gadjah Mada

University, Jl. Fauna No. 3, Bulaksumur, Yogyakarta 55281, Indonesia
Email:yunyer@yahoo.com
2Faculty of Pharmacy, Gadjah Mada University, Jl. Kaliurang Km 4,5 Yogyakarta, 55281
3Integrated Laboratory of Research and Testing, Gadjah Mada University, Jl. Kaliurang Km

4, Yogyakarta 55281

23/11/09 International Seminar in Animal Science and Industry Bogor 23 Nov 2009 1
 INTRODUCTION
qThe falsification of food contents on product
labels is a widespread problem

qFalsifications of meat or food are often

biochemically similar to the materials they
replace, consequently the identification and
measurement extremely difficult.

qHeating of meat may will degrade the DNA

then the isolation of DNA will be difficult
q
2
 INTRODUCTION...........
 Conventional methodology used for the
determination of the species origin in
meat products has been predominantly
based on the immunochemical and
electrophoretic analysis of proteins

Conventionally Weakness Are :

Difficult for species identification
Need long time for protein purification
Unstable by heating and processing
International Seminar in Animal Science and
Industry Bogor 23 Nov 2009
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 OBJECTIVE OF THE RESEARCH
 In the present work we developed a
method for the specific identification of
Pig meats in meatballs based on PCR-
RFLP analysis of a conserved fragment
from the mitochondrial Cytochrome b
gene.
 To know the level of pork contamination in
meatballs which can be detected by
PCR-RFLP

International Seminar in Animal Science and

Industry Bogor 23 Nov 2009

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 Methodology OF THE
RESEARCH

 Sample preparation and DNA

Extraction.
 PCR Amplification
 Enzymatic Digestion of the Amplified
DNA Fragments

International Seminar in Animal Science and

Industry Bogor 23 Nov 2009
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 PCR AMPLIFICATION of Cytochrome b gene

International Seminar on Animal Science and

23/11/09 6
Industry Bogor 23 Nov 2009
  PCR - a copying machine for DNA
molecules
 The purpose of a PCR (Polymerase
Chain Reaction) is to make a
huge number of copies of a gene


2/11/09 Pengantar Bioteknologi 7

 DNA DIGESTION USING RESTRICTION ENZYME
At 55 0C For 3 h

BseDI

C C N N G G

G G N N C C
CUT

The Figure is not really just image the RE

digestion International Seminar on Animal Science and
23/11/09 8
Industry Bogor 23 Nov 2009
RESULTS
Meatball from various
meat mixture and No
body can Justified which
one contents pork or beef

DIFFICULT TO IDENTIFY WHICH ONE

2/11/09
CONTENTS PORK 9
 By DNA Based Methods

2/11/09 Fig. 2 DNA EXTRACTION 10

 The three steps of
PCR

PCR products of
cytochrome b gene
fragments 359 bp long
of samples from
different meatball
products separated by
2% high-resolution
agarosegel
electrophoresis. The
same ingridients with
Fig. 2

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 T h e R E D ig e stio n

BseDI restriction
enzyme on 359 bp
amplicons of
cytochrome b gene
from different
meatball products
separated by 2%
high-resolution
agarose gel
electrophoresis.

PIG MEAT B EEF C h icke n

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 CONCLUSIONS

Our results allow us to conclude that PCR-

RFLP of the mitochondrial Cytochrome b
gene is a suitable alternative that can be
applied to the detection of pig species present
in commercialized products such as
meatballs.

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 THANK YOU

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The homology of
Intraspecies
around 98%

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2/11/09 Pengantar Bioteknologi 16