Preparative in situ hybridization: selection of chromosome region-specific libraries on mitotic chromosomes.
Preparative in situ hybridization: selection of chromosome region-specific libraries on mitotic chromosomes.
复制标题
制备性原位杂交：选择有丝分裂染色体上的染色体区域特异性文库。
DOI：
10.1006/geno.1994.1092
复制
发表时间：
1994
期刊：
影响因子：
4.4
通讯作者：
Davis,L
中科院分区：
文献类型：
--
作者：
Hozier,J;Graham,R;Westfall,T;Siebert,P;Davis,L
我们已经开发了复杂的DNA群体有丝分裂染色体的原位杂交（Prep-ISH）作为产生染色体区域特异性DNA亚群的一种手段。Prep-ISH是两种细胞遗传学技术的组合：DNA分子与有丝分裂染色体的原位杂交和染色体显微切割。在这里，我们目前的测试案例证明这种方法在小鼠和人类基因组的可行性，使用单核，单染色体，或单染色体亚区，以评估灵敏度，特异性和代表性的Prep-ISH技术。Prep-ISH在基因表达和基因组结构的研究中有许多应用，包括组织特异性cDNA和基因组DNA文库的有效细胞遗传学分选。此外，Prep-ISH可能会大大减少候选基因的数量，以帮助基因发现工作，并通过定义的细胞遗传学区域提高开发转录图谱和YAC和粘粒重叠群的效率。
We have developed preparativein situhybridization (Prep-ISH) of complex DNA populations to mitotic chromosomes as a means of generating chromosome region-specific DNA subpopulations. Prep-ISH is a combination of two cytogenetic techniques:in situhybridization of DNA molecules to mitotic chromosomes and chromosome microdissection. Here, we present test cases demonstrating the feasibility of this approach on mouse and human genomes, using single nuclei, single chromosomes, or single chromosomal subregions to assess sensitivity, specificity, and representation of the Prep-ISH technique. Prep-ISH has a number of applications in studies of gene expression and genome organization, including efficient cytogenetic sorting of tissue-specific cDNAs and genomic DNA libraries. In addition, Prep-ISH is likely to dramatically reduce the number of candidate genes to aid in gene discovery efforts and to improve efficiency of developing transcription maps and YAC and cosmid contigs through defined cytogenetic regions.