Lebedev Alexandre
TriLink BioTechnologies, Inc, San Diego, California, USA.
Curr Protoc Nucleic Acid Chem. 2009 Sep;Chapter 4:Unit 4.35 1-17. doi: 10.1002/0471142700.nc0435s38.
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.
2'-脱氧核苷-3'-O-(4-氧代十四烷-1-基)亚磷酰胺（OXT亚磷酰胺）用于制备在3'-末端和倒数第二个核苷酸间连接处以热可裂解的OXT磷酸三酯保护基团修饰的寡脱氧核糖核苷酸引物。与标准DNA引物相比，OXT修饰的引物通过大幅减少或消除PCR假象（如二聚化引物和错配扩增子）的积累，显著提高了聚合酶链反应（PCR）的性能。本文描述了合成OXT修饰的寡核苷酸引物和进行热启动PCR的基本方案。
