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多路连接依赖探针扩增(MLPA)检测拷贝数和突变检测
AlleleID是设计MLPA合成探针或者由MRC Holland引入的多重连接依赖探针扩增技术的程序,当合适的工具包不可用时,研究人员能够扩展这项技术。这是一种快速增长的高通量基因图谱分析方法。可以检测外显子缺失、拷贝数变化和CpG甲基化模式。具有成本效益合适、敏感性高、特好和可再生性好等特点。
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Designing a cross species assay involves the use of a multiple sequence alignment tool to identify conserved stretches of DNA and the design of a hybridization probe for detecting it. Recently there have been advances in micro-organism identification at the taxonomical level using molecular level identification techniques such as real time PCR and microarrays.
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SYBR® Green is the most widely used double-strand DNA-specific dye reported for real time PCR. SYBR® Green binds to the minor groove of the DNA double helix. In the solution , the unbound dye exhibits very little fluorescence. This fluorescence is substantially enhanced when the dye is bound to double stranded DNA. SYBR® Green remains stable under PCR conditions and the optical filter of the thermocycler can be affixed to harmonize the excitation and emission wavelengths. Ethidium bromide can also be used for detection but its carcinogenic nature renders its use restrictive.
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Species Identification: Minimal probes with Optimal primers
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