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Updated: May 5, 2026

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies
Published on: May 22, 2012
Online exercise for the design and simulation of PCR and PCR-RFLP experiments
Rosario María San Millán, Ilargi Martínez-Ballesteros, Aitor Rementeria
1Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006, Vitoria-Gasteiz, Spain. Joseba.bikandi@ehu.es.
Background:
Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism of PCR products (PCR-RFLP) are extensively used molecular biology techniques. An exercise for the design and simulation of PCR and PCR-RFLP experiments will be a useful educational tool.
Findings:
An online PCR and PCR-RFLP exercise has been create that requires users to find the target genes, compare them, design primers, search for restriction endonucleases, and finally to simulate the experiment. Each user of the service is randomly assigned a gene from Escherichia coli; to complete the exercise, users must design an experiment capable of distinguishing among E. coli strains. By applying the experimental procedure to all completely sequenced E. coli, a basic understanding of strain comparison and clustering can also be acquired. Comparison of results obtained in different experiments is also very instructive.
Conclusions:
The exercise is freely available at http://insilico.ehu.es/edu.
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