Peptide nucleic acids (PNAs) have now existed for slightly more than ten years, with interest in and applications of this pseudopeptide DNA mimic steadily increasing during this period. PNAs have rapidly attracted the attention of scientists from a diversity of fields ranging from (bio)organic and biophysical chemistry to prebiotic evolution, and from molecular biology to genetic diagnostics and drug development. Many applications take advantage of the unique properties of PNA—an uncharged pseudopeptide—that distinguish this DNA mimic from more traditional DNA analogs. Rather than trying to create a comprehensive collection of all published methods and protocols involving PNA, many of which have not yet been validated, I have decided to concentrate on select protocols that are either very well established by several groups around the world, such as PCR-clamping and sit.