Content: Extraction of nucleic acids of fungal and bacterial cells. Amplification of the microbial DNA barcoding regions through the polymerase chain reaction (PCR). Detection of specific amplicons through electrophoresis in agarose gel. Sequencing and comparing sequences in the databases to identify genus and species.

Overview: The molecular diagnosis is mainly based on two major cornerstones: nucleic acid hybridization and the amplification methods of these.

Specific objectives:

– Use a commercial system to lyse fungal and bacterial cells and extract the deoxyribonucleic acid (DNA) of both.

– Use the polymerase chain reaction technique (PCR) to amplify regions of the DNA that contain identifying sequences of the species.

– Compare the sequences obtained with those contained in the databases to try and get a higher coefficient of coincidence, to identify the species.

Carrying out the practical:

Each lab bench has yeast (practical 6) and bacteria to be tested (practical 8). The even number lab benches will use the yeast and the odd number lab benches will use the corresponding bacteria.

Microorganisms: Bacteria, Fungi, Virus, ….	Extract genetic material: DNA or RNA	Amplification of the DNA through a PCR

Visualization in agarose gel	Sequencing of the amplicon	Comparison of sequences with the databases

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