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[采用原位杂交的选择性染色体描绘]

[Selective chromosome painting using in situ hybridization].

作者信息

Pérez Losada A, Woessner S, Solé F, Caballín M R, Florensa L

机构信息

Lab. Citología Hematológia, Hospital Central La Alianza, Universidad de Barcelona.

出版信息

Sangre (Barc). 1993 Apr;38(2):151-4.

PMID:8516730
Abstract

A method of in situ hybridization with entire chromosome-specific DNA libraries for visualizing individual human chromosomes has been developed and applied to the detection of structural aberrations in both metaphase and interphase cells. Unlabeled human genomic DNA is used to inhibit the cross-hybridization of repetitive sequences in the library that bind to multiple chromosomes. The remaining single-stranded DNA is hybridized to specimens of interest and detected with fluorescent or enzyme labeled biotin conjugates following post-hybridization washes. This general approach is called "chromosome painting" or "chromosomal in situ suppression (CISS)" hybridization. In the present report, DNA inserts from recombinant libraries from chromosomes 1, 4, and 9 has been applied on controls and patients in order to decorate specifically their complementary chromosomes. Numerical changes, deletions and chromosomal translocations involving these chromosomes can be strikingly visualized.

摘要

一种利用全染色体特异性DNA文库进行原位杂交以可视化单个人类染色体的方法已被开发出来,并应用于检测中期和间期细胞中的结构畸变。未标记的人类基因组DNA用于抑制文库中与多条染色体结合的重复序列的交叉杂交。剩余的单链DNA与感兴趣的标本杂交,并在杂交后洗涤后用荧光或酶标记的生物素缀合物进行检测。这种通用方法被称为“染色体描绘”或“染色体原位抑制(CISS)”杂交。在本报告中,来自1号、4号和9号染色体重组文库的DNA插入片段已应用于对照和患者,以便特异性地标记其互补染色体。涉及这些染色体的数目变化、缺失和染色体易位可以显著地可视化。

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