ABSTRACT
Inverse PCR1-3 is a useful technique for supplementing conventional PCR screens that often yield only a substantial fragment of the gene of interest. An inverse PCR strategy allows the user to “walk” both upstream and downstream of a known DNA sequence in order to obtain additional sequence and facilitates amplification of unknown DNA flanking sequences without the labor involved in constructing and screening libraries. The main steps are as follows (Figure 31.1):
1. Purify and cut the genomic DNA sample with appropriate restriction enzymes (EcoRl and Hind III in this example). Ligate the fragments into circular templates.
