How to Test for Animal Speciation

How to Test for Animal Speciation

The 2013 horse meat scandal in Europe has made the testing for meat species important. Besides wrong labelling of meat, meat speciation is important for halal and kosher products where pork is required to be absent. Such meat adulterations can be detected using real-time PCR and can be quantified. Besides meat speciation, Bioscience Diagnostics Singapore provides kits for fish and milk adulteration. Identification of meat at the specie level is done by analyzing the product for the specific DNA using real-time PCR. However sub specie identification is not possible due to the absence of any highly conserved region for the sub specie. Raw, processed, cooked meat speciation is possible as long as about 10 copies of DNA is present.

The steps involved in meat speciation are:

1. Sampling

4) Pic 2_Sampling

Meat sample has: -

Nucleus – single copy DNA

Mitochondria – multi copy DNA

DNA is a cellular blueprint and contains information on its own species in highly conserve regions.

2. Getting to the DNA

4)-Pic-3_Getting-to-the-DNA-1

Cells have a cell wall which protects the DNA. To get to the DNA, the cell has to be broken with cell lysis. This is done with the following reagents.

  • 400µl lysis buffer
  • 40 µl proteinase K
  • React for 30mins at 52oC
  • 200µl binding buffer
4)-Pic-4_Getting-to-the-DNA-2

After lysis, cell has many debris of which will inhibit extraction of DNA.
Purification via spin filter with DNA trapped.
Wash twice with 550µl wash buffer.

4)-Pic-4_Getting-to-the-DNA-3

Finally release trapped DNA with wash buffer.

4) Pic 6_Getting to the DNA 4

Multiply DNA using prepared kit and multicycler.

4) Pic 7_Getting to the DNA 5

Get Results from software

For details on this kit and others contact Bioscience Diagnostics Singapore sales.

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