Long PCR Methodology

In this chapter, we detail protocols of long polymerase chain reaction (PCR) and long RT-PCR, which we have found to be versatile, sensitive, and straightforward to optimize. We have used these protocols with success on several different templates, including lambda phage DNA, HAV, HBV, HCV (1 ), torovirus (2 ), coxsackie B6 virus (3 ), and human beta galactosidase mRNA (R. Tellier, unpublished data). The guanine-cytosine (GC) content of these templates varied from 37.8 to 58.8%. These protocols have also been used on genomic human DNA with success (4 ).