Pcr Analysis Of Minced Meat Sample

Analysis of minced meat samples using Polymerase Chain Reaction (PCR) and Gel Electrophoresis to investigate adulteration in the product

Food Traceability and Genomics

John Barry Title: Analysis of minced meat samples using Polymerase Chain Reaction (PCR) and Gel Electrophoresis to investigate adulteration in the product

Name: John Barry

Date: 7/10/14

Aim: The aim of the experiment was to examine minced meat samples for adulteration by amplifying extracted DNA using a PCR method. Gel Electrophoresis was then carried out on the PCR product to separate the DNA molecules.

Introduction: Polymerase Chain Reaction (PCR), a technique commonly used in research, diagnostics and the detection of particular genes , was developed by Kary

The Food Safety Authority of Ireland (FSAI) defines food fraud as “the act of deliberately and illegally placing food on the market with the intention of deceiving the customer, usually for financial gain” (FSAI, 2013). While it would be hoped that the systems in place within the food supply chain (traceability, certification etc.) would prevent food fraud , the economic gain associated with its practise is so great there will always be a risk of its occurrence. The associated economic gain can be through tax and customs duty evasions or through the use of cheaper, alternative substances. In the case of the horse meat crisis the economic gain was through the use of an alternative substance. In the horse meat case there was no significant health risk to those who had consumed the product, however consumer confidence in processed meat products was damaged and more importantly Irelands international image as a producer of quality food was brought into

The comb was then placed into the solution to create the wells. Once the gel had solidified (4-5 minutes) the comb was carefully removed. The solidified gel was then placed into the buffer chamber ensuring that the wells were located closest to the negative terminal. This ensures that the DNA will travel through the gel in the direction of the positive terminal. If the wells are placed closest to the positive terminal it will result in the DNA running off the gel and a failed procedure.

1 X TAE buffer (quantity unknown) was added to the buffer chamber until the gel was completely submerged. The buffer solution was added to create the optimum pH and facilitate electrical conductivity within the chamber. Loading dye (5 µl) was then added to the first well using a carefully pipetting action. PCR product (5 µl) was added to each well and loading dye (5 µl) was added to the final well. The PCR chamber was then sealed and a current of 110mA applied for one hour.

The solidified gel was then removed from the chamber and placed under UV light. A digital image was then taken, allowing for analysis to be carried