Gosden J, Lawson D
MRC Human Genetics Unit, Western General Hospital, Edinburgh, UK.
Hum Mol Genet. 1994 Jun;3(6):931-6. doi: 10.1093/hmg/3.6.931.
We describe a method for rapid identification of chromosomes at metaphase, and quantification of chromosomes in interphase, by annealing oligonucleotide primers, derived from chromosome-specific subsets of repeated DNA families, to the DNA of cytological preparations, and enzymatic extension with the incorporation of labelled nucleotides. The method is equally applicable to normal cells or those from somatic cell hybrids. Where the labelled product is too small or of too low a copy number to be readily seen after the single extension reaction, we have developed a method for cyclic amplification of the labelled DNA, enabling clear visualization of the signal.
我们描述了一种方法,通过将源自重复DNA家族的染色体特异性亚组的寡核苷酸引物与细胞学制剂的DNA退火,并结合标记核苷酸进行酶促延伸,来快速鉴定中期染色体并定量间期染色体。该方法同样适用于正常细胞或体细胞杂种细胞。当标记产物太小或拷贝数太低以至于在单次延伸反应后不易观察到时,我们开发了一种标记DNA的循环扩增方法,从而能够清晰地观察到信号。