The aim of this study is to establish a HRM (high resolution melting curve) method for detection of deletion in human BIM gene and to detect this site deletion with the above method in 30 lung cancer samples and 30 normal samples.
Methods
The primers for detection of BIM deletion were designed and synthesized. The HRM method for geng deletion was established. And select the part of samples to detect BIM delection by normal PCR and sequencing assay. The Tm value of wild type PCR products was higher than that of the deletion PCR products. The difference of the corresponding Tm value is 2.5 ℃.
Results
By detection with HRM methods, 1 samples were confirmed to be mutant, 7 samples were confirmed to be heterozygous and the other 22 samples were all wild type in the lung cancer samples. 2 samples were confirmed to be heterozygous and the other 28 samples were all wild type in the normal samples.
Conclusion
The HRM method for detection of BIM deletion established in this study is a sensitive, accurate, simple and high throughput method.
The melting temperature of wild-type is higher than the deletion. Ater the data processing the significant difference of the melting curve peak temperature is about 2.5 ℃ (——wild type——deletion).
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