The production of PCR products with 5' single-stranded tails using primers that incorporate novel phosphoramidite intermediates.
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.
Newton
C R
CR
ZENECA Pharmaceuticals, Alderley Park, Macclesfield, Cheshire, UK.
Holland
D
D
Heptinstall
L E
LE
Hodgson
I
I
Edge
M D
MD
Markham
A F
AF
McLean
M J
MJ
D000469:Alkaline Phosphatase / Q000378:metabolism; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009838:Oligodeoxyribonucleotides; D009943:Organophosphorus Compounds / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*
D004277:DNA, Single-Stranded; D009838:Oligodeoxyribonucleotides; D009943:Organophosphorus Compounds; D000469:Alkaline Phosphatase