DEVELOP GBA BASED MTDNA & Y CHROMOSOME HAPLOTYPE SYSTEM

描述（改编自研究者摘要）：法医分析 将基于非假常染色体区域的分析开发系统 和线粒体高变区 基因组其目的是增加歧视的可能性， 在性侵犯案件和大规模灾难中收集的DNA证据 网站.该方法将基于检测单核苷酸 通过遗传位作图（GBA）技术检测SNP， 已经被Molecular Tool（最近收购的GeneScreen Inc）使用。 所提出的用于鉴定SNP的技术已经被设计用于高 吞吐量分析 它包括以下分析步骤1。 PCR 使用常规引物和硫代磷酸化引物对感兴趣区域进行扩增。 2. 使用T7外切核酸酶（未保护）制备硫代磷酸化ssDNA 链被消化）。 3. 将ssDNA捕获到固定的引物（两个 平行平板以检测所有四种碱基）。 4. 单碱基 用标记有双脱氧核苷三磷酸的引物延伸 生物素或荧光素。 5. 通过检测延伸产物， 使用碱性磷酸酶标记抗- 荧光素或过氧化物酶标记的荧光素。 结合型碱性磷酸酶 首先检测，然后洗涤孔， 双色检测（Two Color Detection） 最初，该方法将在宏观尺度上使用96孔 微量滴定板和比色检测。 随后， 微型化到载玻片上，并采用直接荧光检测， 捕获的酶标记的缀合物，最终使用四种不同的 荧光标记，使得每个SNP基因座处的所有四个核苷酸都可以被标记。 立即询问。 拟议商业应用：不可用

DESCRIPTION (Adapted from investigator's abstract): A forensic analysis system will be developed based on analysis of non-pseudoautosomal regions of the Y chromosome and the hypervariable region of the mitochondrial genome. The intent is to increase the probability of discrimination for DNA evidence collected in sexual assault cases and from mass disaster sites. The methodology will be based on the detection of single nucleotide polymorphisms (SNP) by genetic bit mapping (GBA) techniques that are already used by Molecular Tool (a recent acquisition of GeneScreen Inc). The proposed technology for identifying SNPs has been designed for high throughput analysis. It involves the following analytical steps 1. PCR of region of interest using a regular and a phosphorothioated primer. 2. Preparation of phosphorothioated ssDNA using a T7 exonuclease (unprotected strand is digested). 3. Capture of ssDNA to immobilized primers (two plates in parallel for detection of all four bases). 4. Single base extension of the primer with a dideoxynucleoside triphosphate labeled with biotin or fluorescein. 5. Detection of the extension product via the biotin or fluorescein tag using alkaline phosphatase-labeled anti- fluorescein or peroxidase labeled antibiotin. Bound alkaline phosphatase is detected first and then the well is washed and any bound peroxidase is measured (two color detection). Initially, the method will be developed on a macroscale using 96-well microtiter plates and colorimetric detection. Subsequently, it will be miniaturized onto glass slides and employ direct fluorescent detection of captured enzyme labeled conjugate, ultimately using four different fluorescent labels so that all four nucleotides at each SNP locus can be interrogated at once. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE