通过双变量流式细胞分选，随后进行简并寡核苷酸引物PCR，分离大鼠染色体特异性涂染探针。

Isolation of rat chromosome-specific paint probes by bivariate flow sorting followed by degenerate oligonucleotide primed-PCR.

作者信息

Hoebee B, de Stoppelaar J M, Suijkerbuijk R F, Monard S

机构信息

Laboratory of Carcinogenesis and Mutagenesis, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.

出版信息

Cytogenet Cell Genet. 1994;66(4):277-82. doi: 10.1159/000133712.

DOI:10.1159/000133712

PMID:8162707

Abstract

The flow karyotype of rat chromosomes was determined by dual beam flow cytometry. Eighteen fractions were sorted and subsequently amplified by degenerate oligonucleotide primed-PCR as described by Telenius et al. (1992a, 1992b). The PCR products were labeled and used as probes for fluorescence in situ hybridization on rat fibroblast metaphases. The amplified chromosomes were detectable as bright chromosome paints and in most cases the signal was evenly distributed along the whole chromosome except for the centromeric region in half of the chromosomes. The results show that chromosomes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 19, 20, X and Y can be sorted as individual fractions, whereas chromosomes 11, 13, 14, 15 and chromosomes 16, 17 and 18 are clustered together in the flow karyotype.

摘要

通过双光束流式细胞术确定了大鼠染色体的流式核型。分选了18个组分，随后按照Telenius等人（1992a，1992b）所述，通过简并寡核苷酸引物PCR进行扩增。PCR产物被标记，并用作大鼠成纤维细胞中期荧光原位杂交的探针。扩增的染色体可作为明亮的染色体涂染物被检测到，并且在大多数情况下，信号除了在一半染色体的着丝粒区域外，沿整个染色体均匀分布。结果表明，染色体1、2、3、4、5、6、7、8、9、10、12、19、20、X和Y可以作为单个组分进行分选，而染色体11、13、14、15以及染色体16、17和18在流式核型中聚集在一起。