DNA amplification and multiplexing
After a brief reminder of the principles underlying the polymerase chain reaction (PCR), Chapter 6 “DNA amplification and multiplexing” discusses the choice of a DNA polymerase for the PCR. In particular, it warns against the use of proofreading polymerases, that can lead to a substantial loss of PCR specificity. Chapter 6 insists on the benefits of including different types of controls in the PCR (e.g., PCR negatives and positives, tagging system controls, etc.). The most common causes of PCR failures and their solutions are addressed, as well as the precautions to take to avoid and monitor contaminations. Chapter 6 also deals with the particular case of blocking oligonucleotides, which aim at reducing the amplification of undesired sequences. It gives some valuable guidelines to design such oligonucleotides and use them efficiently. Finally, Chapter 6 presents different strategies for tagging individual samples during the amplification, to allow subsequent multiplexing during the sequencing step.
Genetic differentiation and gene flow between the Tunisian ovine breeds Barbarine and Western thin tail using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis
