Newton C R, Holland D, Heptinstall L E, Hodgson I, Edge M D, Markham A F, McLean M J
ZENECA Pharmaceuticals, Alderley Park, Macclesfield, Cheshire, UK.
Nucleic Acids Res. 1993 Mar 11;21(5):1155-62. doi: 10.1093/nar/21.5.1155.
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.
我们制备了几种新型亚磷酰胺，并合成了内部包含这些亚磷酰胺的寡核苷酸。寡核苷酸模板中这些残基的存在对Taq DNA聚合酶引发的合成造成了不可逾越的障碍。当作为聚合酶链反应产物延伸时，这些寡核苷酸在修饰中间体的插入位点之外不再作为聚合酶的模板，从而在扩增产物上产生单链尾巴。然后这些尾巴可用于固相捕获和PCR产物的比色检测。
