摘要
目的 制备单链探针替代原有的双链探针 ,以解决RDPCR技术中存在的杂交效率较低及特异性较差的问题。方法 应用不对称PCR及单引物PCR技术制备大鼠p5 3基因外显子 7单链探针 ,再经RDPCR扩增比较单链与双链探针的杂交效果。结果 不对称PCR及单引物PCR技术制备单链探针均获得成功 ;目的基因单链探针杂交条带清晰且几乎无背景值。结论 与双链探针比较 ,单链探针的应用可以明显提高RDPCR技术杂交的敏感性及特异性。
Objective\ Preparing single-stranded (ss) probes in place of double- st randed(ds) probes to improve the hybridization efficiency and specificity of randomized terminal linker-dependent PCR(RDPCR). Methods Using asymmetric PCR and single-primer PCR to prep are ss probes of e xtron 7 of p53 gene in rat, then comparing the results hybridized with ss pr obes and ds probes. Results Preparation of ss probes by asymmetric PCR and single-pr imer PCR gets success. Hybridization results showed that the ss probes could get better signals and less noise than ds probes. Conclusion In co mparison with ds probes, the application of ss probes can increase the hybridization sensitivity and specificity of RDPCR.
出处
《卫生研究》
CAS
CSCD
北大核心
2005年第2期172-174,共3页
Journal of Hygiene Research
基金
国家自然科学基金资助项目 (No .30 0 70 648)