title: Heat-activatable primers for hot-start PCR: oligonucleotide synthesis and basic PCR setup.
abstract: 2'-Deoxyribonucleoside-3'-O-(4-oxotetradec-1-yl) phosphoramidites (OXT phosphoramidites) are used to prepare modified oligodeoxyribonucleotide primers containing heat cleavable OXT phosphotriester protecting groups at 3'-ultimate and penultimate internucleotide linkages. The OXT-modified primers significantly improve performance of the polymerase chain reaction (PCR) compared to standard DNA primers by substantially reducing or eliminating the accumulation of PCR artifacts such as dimerized primers and misprimed amplicons. Basic protocols for synthesis of OXT-modified oligonucleotide primers and for performing hot-start PCR are described.
authors.affiliation: TriLink BioTechnologies, Inc, San Diego, California, USA.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Lebedev
mesh_terms: D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D009943:Organophosphorus Compounds / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D009841:Oligonucleotides; D009943:Organophosphorus Compounds; C434331:phosphoramidite