TEMPLATE-DIRECTED INCORPORATION OF NUCLEOTIDE MIXTURES USING AZOLE-NUCLEOBASE ANALOGS

DNA that encodes elements for degenerate replication events by use of artificial nucleobases offers a versatile approach to manipulating seq uences for applications in biotechnology, We have designed a family of artificial nucleobases that are capable of assuming multiple hydrogen bonding orientations through internal bond rotations to provide a mea ns for degenerate molecular recognition, Incorporation of these analog s into a single position of a PCR primer allowed for analysis of their template effects on DNA amplification catalyzed by Thermus aquaticus (Tag) DNA polymerase, All of the nucleobase surrogates have similar sh apes but differ by structural alterations that influence their electro nic character, These subtle distinctions were able to influence the Ta o DNA polymerase dependent incorporation of the four natural deoxyribo nucleotides and thus, significantly expand the molecular design possib ilities for biochemically functional nucleic acid analogs.