A Rapid Method for Generation of Homologous Recombinant Fragments—Fusion PCR
- VernacularTitle:一种高效构建同源重组DNA片段的方法——融合PCR
- Author:
Min LI
;
Qian YANG
;
- Publication Type:Journal Article
- Keywords:
Fusion PCR Recombinant fragment Homologous arm Resistant gene
- From:
China Biotechnology
2006;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Fusion PCR, which employs chimeric primers to generate PCR products with complementary ends in its amplifications, is a rapid and flexible method in joining different DNA fragments. This method can assemble DNA fragments without the treatment of restriction endonucleases and T4 DNA ligase. It offered a shortcut for the construction of homologous recombinant fragments. Through assembling three recombinant fragments successfully, fusion PCR procedure was improved and manipulation essentials of fusion PCR were described in detail. The results indicated that the improved procedure can assemble three or four fragments simultaneously, and the length of each fusion product is above 4.5 kb. The result recombinants were proposed to be use in further experiments, which structures had been confirmed by sequencing.