文献类型： 外文期刊
作者： Zhang, YL 1 ; Zhang, DB 2 ; Li, WQ 2 ; Chen, JQ 2 ; Peng, YF 2 ; Cao, W 2 ;
作者机构： 1.Shanghai Acad Agr Sci, Agrobiotech Res Ctr, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
2.Shanghai Acad Agr Sci, Agrobiotech Res Ctr, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China; Nanjing Univ, Dept Biol Sci & Technol, Nanjing 210093, Peoples R China; GenoMultix Co Ltd, Dept Technol, Shanghai 200233, Peoples R China; Chinese Acad Agr Sci, Inst Crop Protect, Beijing 100094, Peoples R China
期刊名称：NUCLEIC ACIDS RESEARCH （ 影响因子：16.971； 五年影响因子：15.542 ）
ISSN： 0305-1048
年卷期： 2003 年 31 卷 20 期
页码：
收录情况： SCI
摘要： A novel real-time quantitative polymerase chain reaction (PCR) method using an attached universal template (UT) probe is described. The UT is an approximately 20 base attachment to the 5' end of a PCR primer, and it can hybridize with a complementary TaqMan probe. One of the advantages of this method is that different target DNA sequences can be detected employing the same UT probe, which substantially reduces the cost of real-time PCR set-up. In addition, this method could be used for simultaneous detection using a 6-carboxy-fluorescein-labeled UT probe for the target gene and a 5-hexachloro-fluorescein-labeled UT probe for the reference gene in a multiplex reaction. Moreover, the requirement of target DNA length for UT-PCR analysis is relatively flexible, and it could be as short as 56 bp in this report, suggesting the possibility of detecting target DNA from partially degraded samples. The UT-PCR system with degenerate primers could also be designed to screen homologous genes. Taken together, our results suggest that the UT-PCR technique is efficient, reliable, inexpensive and less labor-intensive for quantitative PCR analysis.
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