Multiplex Pcr Procedure

1 taq polymerase buffer 4 mm mgcl 2 300 nm concentrations of each of the primers listed in table table2 2 400 μm concentrations of deoxynucleoside triphosphates 5 u of taq polymerase and 50 ng of staphylococcal dna.

Multiplex pcr procedure.

This process amplifies dna in samples using multiple primers and a temperature mediated dna polymerase in a thermal. Multiplex pcr introduction of multiplex pcr. A multiplex pcr procedure that detects six major virulence genes flic stx1 stx2 eae rfbe and hlya in escherichia coli o157 h7 was developed. In a multiplexing assay more than one target sequence can be amplified by using multiple primer pairs in a reaction mixture.

The multiplex pcr was performed in a 50 μl volume with the gibco brl taq dna polymerase system life technologies inc rockville md containing the following. Slideshare uses cookies to improve functionality and performance and to provide you with relevant advertising. References multiplex pcr. Multiplex pcr is a variant of pcr method in which more than one target sequence is amplified using multiple sets of primers within a single pcr mixture.

D pcr buffer concentration. More than two sets of primers amplify several different sequences of the templet dna or sequences of multiple temples in a single pcr reaction is known as multiplex pcr in a simple language we can say it is a combination of different pcr reaction thus more reagents are needed to amplify various template regions. Multiplex pcr was performed with mixture y3 in 1 4 pcr buffer using pcr program e and gradually raising the concentration of mgcl 2. Analyses of the available sequences of the six major virulence genes and the published primers allowed us to develop the six gene multiplex pcr protocol that maintained the specificity of each primer pair.

This enables amplification of several gene segments at the same time instead of specific test runs for each. What is a multiplex pcr.