Zheng W, Fu M, Wang X
Cancer Institute, CAMS/PUMC, Beijing.
Zhonghua Yi Xue Za Zhi. 1996 Apr;76(4):305-7.
To localize the differentiation-relevant cDNA RA538 (3.8kb) onto the human chromosome 8q2.
A modified non-isotopic labeling technique was used.
The cDNA RA538 was obtained by subtraction hybridization from human esophageal cancer cell line EC8712 induced by retinoic acid. Fluorescence in situ hybridization (FISH) using double-labeled probes showed clear and paired hybridization signals at the corresponding regions of both two chromatids in 18 out of 60 metaphases examined. While by single-labeling, only 7 were positive in 60 metaphases observed and the fluorescent spots were seen only at one chromatid.
The modified FISH protocol is useful for mapping cDNA sequences of a few kilobases.
将与分化相关的cDNA RA538（3.8kb）定位到人类染色体8q2上。
采用改良的非同位素标记技术。
通过对维甲酸诱导的人食管癌细胞系EC8712进行消减杂交获得cDNA RA538。使用双标记探针的荧光原位杂交（FISH）显示，在60个检测的中期相中，有18个中期相的两条染色单体的相应区域出现清晰且配对的杂交信号。而单标记时，在观察的60个中期相中只有7个呈阳性，且荧光点仅出现在一条染色单体上。
改良的FISH方案有助于绘制几千碱基的cDNA序列图谱。
