PCR amplification of nucleic acids is a fundamental technique used in many molecular biology  laboratories. Despite its wide use, certain GC-rich regions of DNA, such as mycobacterial  disease targets, still remain a challenge for amplification. Sequences high in GC content are  associated with the formation of secondary structure, which prevents adequate strand separation  and DNA polymerase amplification. As a consequence, mispriming is prominent, complicating  specific product formation.