ABSTRACT

The ability to determine nucleic acid sequences is of utmost importance for the detailed study of biological systems. Pyrosequencing− is the first alternative to the traditional Sanger method for de novo DNA sequencing.1,2 The Pyrosequencing technology is especially suited for short-read sequencing,3 although the method has recently been improved to allow sequencing of up to hundred bases.4 It is broadly applicable for analysis of single nucleotide polymorphism (SNPs)5 ç8

and for identification of short DNA sequences used in bacterial,9 fungal (unpublished data), and viral typing.10 In addition, the methodological performance of the Pyrosequencing technology has been used to determine difficult secondary structures,11 mutations,12,13 and for tag sequencing of a selected cDNA library.14