The production of PCR products with 5' single-stranded tails using primers that incorporate novel phosphoramidite intermediates.

AUTOR(ES)

Newton, C R

RESUMO

We have prepared several novel phosphoramidites and have synthesised oligonucleotides incorporating them internally. The presence of these residues in an oligonucleotide template presents an impossible barrier to primed synthesis by Taq DNA polymerase. When extended as polymerase chain reaction products, these oligonucleotides no longer serve as templates for the polymerase beyond the insertion sites of the modified intermediates, thereby producing single-stranded tails on amplification products. These tails can then be used for solid phase capture and colorimetric detection of PCR products.

Documentos Relacionados

• Single-stranded replication intermediates of ribosomal DNA replicons of pea.
• Direct sequencing of double-stranded DNA PCR products via removing the complementary strand with single-stranded DNA of an M13 clone.
• The RecOR proteins modulate RecA protein function at 5′ ends of single-stranded DNA
• Subtractive hybridization system using single-stranded phagemids with directional inserts.
• Studies on nucleic acid reassociation kinetics: reactivity of single-stranded tails in DNA-DNA renaturation.