A fluorescence polarization-based method to detect BRCA2 mutation
AIM: To develop a rapid to detect BRCA2 gene mutation for large-scale screening. METHODS: The DNA fragment covering the mutation site was amplified with PCR method. The fluorescents labeled terminator (R110-acyGTP or TAMRA-acy TTP) was incorporated to the 3'-end of detecting probe via template-directed dye-terminator incorporation ( TDI) reaction. The mutation was detected by fluorescence polarization (FP) measurement. RESULTS: The new method has been established based on the TDI-FP platform. The concentration of primers and dNTPs and the number of PCR cycle have been optimized. To evaluate this method, genomic DNA samples from breast cancer patients and healthy controls were analyzed. The results were perfectly consistent with those determined by direct sequencing. CONCLUSION: This new method enables the high throughput, accurate and rapid detection of BRCA2 mutation. It can be used to large-scale screening in clinic laboratories.
NETL
NSTL
万方数据
维普
