Thermal-bias PCR: generation of amplicon libraries without degenerate primer interference.

ABSTRACT: The polymerase chain reaction (PCR) has been used to amplify specific gene regions for many taxonomic studies and there have been substantial efforts to develop protocols that efficiently amplify target regions from a majority of mixed-template populations. Most protocols include the use of degenerate oligonucleotide primer pools, which contain mixed nucleotide sequences to improve priming from templates containing non-consensus sequence variations in their primer-binding sites. In this work, computational modeling and experimental measurements revealed that degenerate primers reduce efficiency well before a substantial product pool has been generated. It was also discovered that non-degenerate primers produced amplicons significantly better than their degenerate counterparts when amplifyi

SUBMITTER: Moore SD 

PROVIDER: S-EPMC12558157 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature