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A simple method for PCR based analyses of immunohistochemically stained, microdissected, formalin fixed, paraffin wax embedded material.

机译:一种简单的基于PCR的免疫组织化学染色,显微解剖,福尔马林固定,石蜡包埋材料的分析方法。

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摘要

Microdissection was performed on sections cut from formalin fixed, paraffin wax embedded archival material, which had been subjected to conventional immunohistochemistry. Crude DNA extracts, which were obtained from these microdissected samples by a simple microwave step, were then added directly to amplification reactions. Analyses using a range of polymerase chain reaction (PCR) based techniques, including microsatellite repeat polymorphism analysis at the NM23-H1 locus and sequencing of exons 5, 7, and 8 of the p53 gene, were performed successfully. Universal PCR amplification was also carried out on the microdissected material and probes suitable for use in comparative genomic hybridisation (CGH) were obtained in all cases. This technique will enable a range of effective genetic analyses to be carried out on specific subsets of cells that have been characterised previously by immunohistochemistry.
机译:从福尔马林固定的,石蜡包埋的档案材料上切下的切片进行了显微解剖,该切片已进行了常规免疫组织化学分析。通过简单的微波步骤从这些显微切割的样品中获得的粗制DNA提取物随后直接添加到扩增反应中。使用一系列基于聚合酶链反应(PCR)的技术成功进行了分析,包括在NM23-H1基因座上的微卫星重复多态性分析和p53基因的外显子5、7和8的测序。还对显微切割后的材料进行了通用PCR扩增,在所有情况下均获得了适用于比较基因组杂交(CGH)的探针。这项技术将能够对先前已通过免疫组织化学表征的特定细胞亚群进行一系列有效的遗传分析。

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