How to Optimize Multiplex PCR for Histology Applications?
Successful optimization of multiplex PCR involves several steps:
Primer Design: Use software tools to design primers that have similar melting temperatures (Tm) and do not form secondary structures or dimers. Reaction Conditions: Optimize key parameters such as annealing temperature, magnesium ion concentration, and DNA polymerase choice. Template Quality: Ensure high-quality DNA or RNA extraction from histological samples to avoid degradation and contamination. Controls: Include positive and negative controls to validate the specificity and efficiency of the multiplex PCR.
