Chromosome mapping of the human cytidine-5?-triphosphate synthetase (CTPS) gene to band 1p34.1?p34.3 by fluorescence in situ hybridization

Abstract

The human cytidine-5'-triphosphate synthetase (CTPS) gene was mapped by a direct mapping system combined with fluorescence in situ hybridization and replicated prometaphase R-bands. By high-resolution banding analysis, the signals were localized to band 34.1-34.3 of the short arm of chromosome 1; 1p34.1-p34.3. Simple procedures for the detection of R-bands are described.

Precise mapping of the EGF receptor gene on the human chromosome 7p12 using an improved fish technique

“…For example, G-banding shows no fluorescence and therefore the combination of G-banded chromosomes with FISH signals is not feasible. R-banding has been successfully used (Takahashi et al, 1991) FITC probe signals can not be detected because both show green fluorescence excited by the same wave length. Texas Red (TR) has been used successfully (Fukuyama and Shimizu,I992) and similar fluorescent dyes like rhodamine can be used.…”

High-resolution chromosome R-banding in lymphoblastoid cell lines by the combined use of cell synchronization and ethidium bromide treatment

“…At appropriate time intervals, the cells were harvested for chromosome preparation following short-term treatments with either Colcemid (0.02/~g/ml, 30 min) alone or Colcemid (0.02/zg/ml, 30 min) and EB (7.5/~g/ml, 1.5 hr) in combination. R-banded chromosomes were prepared as reported previously (Takahashi et al, 1991) with slight modifications: the chromosome slides made using the above procedures were stained with Hoechst 33258 (1/lg/ml, 10 min) and mounted in 2• (pH 7.4) under cover slips. They were then placed on a 75~ hot plate for 9 min while being exposed to UV light at a distance of 1.5 cm for the last 6 rain, and stained with Giemsa.…”

Identification of iso(18p) marker chromosome by fluorescencein situ hybridization with single-copy DNA probe

“…To induce R-banding stimulated PBLs were synchronized by blocking the cell cycle for 16 h with excess thymidine, and were treated with 5-bromo-2'-deoxyuridine (BrdU) during the final late S-phase before harvesting (Viegus-Pequignot and Dutrillaux, 1978). For R-banding and FISH analysis, chromosome spreads were stained with Hoechst 33258 and were exposed to UV light for 10 rain (Takahashi et al, 1991). To induce multiple 4',6-diamino-2-phenylindole (DAPI)-banding, denaturation time was shortened to 30-60 sec as described (Heng and Tsui, 1993), and the step of UV exposure was omitted.…”

“…FISH was performed according to the methods described elsewhere (Lichter et al, 1988;Takahashi et al, 1991). Cosmid DNAs were labeled by nick translation using the enzyme kit (Boehringer Mannheim, GmbH) with biotin-ll-dUTP (Boehringer Mannheim) and hybridized to chromosome DNA under conditions that suppress signals from repetitive sequences (Lichter et aL, 1988).…”