Effect of Meat Processing on

Genomic DNA Quality and

Specific Gene Amplification

Valdy azhari/ 155100500111011


Introduction
The background of the researcher
doing the research and the
purpose of the research
Presentation Outline
Material and Method
DNA Extraction from the minced
fresh/processed meat (Chicken
meat & Pork) and Amplification of
chicken meat and pork DNA that
have been extracted by PCR
Result and Discussion
The result of the research from
identifying the effect of cooking
and processing on quality of
extracted DNA and the conclusion
of the research
Introduction
Research Background

Is it Adulterated meat?
Meat need to be identified
Research Background

PCR technique can identify

Is Gene from cooked species differentiation
meat can be detected?
Research Purpose

“ To Evaluate the Effect of Cooking and Processing

on Quality of
Extracted DNA and The Amplification of Specific
Gene, using PCR “
Material and Method
 Meats Minced
Sample (separately)

Chicken Meat
Each 3 250g sample
The raw chicken meat samples were Treated with 3
collected from the sale counter, Central different temperature
Avian Research Institute, Izatnagar

Pork Meat

A
raw pork samples were collected
B C D
autoclaved blocks (15
from local market Oven cooked blocks steam cooked blocks lb pressure, 1200 C, 30
(720C, 30 min) (steam 9O0 C, 30 min) min)
Proccesed meat
Chicken patties and canned chicken
preparation were bought from local
market
 Absolute alcohol

DNA Extraction

Extracted using
Precipitated in high Peleted and washed
phenol-chloroform
salt concentraton with 70% alcohol
method

100g meat
Dissolved in TE buffer

Lambda DNA digested with gel was visualized under DNA resolved on 0.8%
HindIII and EcoR I was used UV transilluminator and agarose gel at 5 V/cm
photographed for 2 h
as molecular size marker.

DNA
Amplification of 12S rRNA
gene
Diluted to the 25 µl reaction Amplification in PCR
Concentration of 25- volume(PCR Mix) used using Girish et al
75 ng/µl for PCR in PCR condition

Resolved on 1,4%
DNA Agarose

PCR Mix PCR Condition

each PCR amplification reaction was set in a
volume of 50 ml with 5 ml of 10X PCR buffer
(100 mM Tris HCl, pH 9.0, 15mM MgCl, 500
mM KCl and 0.1% gelatin), 1ml of10 mM
dNTP mix, 1 ml each of (20 pmoles) forward
and reverse primers, 1U of Taq DNA
polymerase and 50 ng of purified DNA
Were as follows: 5 min at
940 C for initial
denaturation, followed by
30 cycles of amplification
(45 s at 940 C, 45 s at 600 C
and 1 min at 720 C) and
final extension for 10 min at
720 C.
Result and Discussion
Result
Result
• Sherring DNA increased with the severity of
Processing
• little shearing of DNA was observed in meat
cooked at 72C For 30 min considerable shearing
occurred in autoclaved meat
• In chicken patties and canned chicken also got
shearing

• A 450 bp fragment of 12S rRNA gene was successfully

amplified using genomic DNA from raw meat from
chicken and pig also processed meat as template and
universal primers.
• however the intensity of the bands amplified from
autoclaved meat was comparatively less in comparison
to others
• These results indicated that processing of meat does not
affect the amplification of 12S rRNA gene using the
universal primers
 CONCLUSION

• It suggested that though the cooking and or processing affected

the quality of genomic DNA extracted, but it did not affect the
amplification of 12S rRNA gene
• PCR can amplify relatively short sequences in highly degraded
DNA. It was proved by 450 bp fragment of mt 12S rRNA gene
amplification in highly processed meats
Thank You!
ANY QUESTIONS?