ULS: a versatile method of labeling nucleic acids for FISH based on a monofunctional reaction of cisplatin derivatives with guanine moieties.
The broad extension of an existing chemical DNA labeling technique for molecular cytogenetics is described. Called the Universal Linkage System (ULS(TM)), it is based on the capability of monoreactive cisplatin derivatives to react at the N7 position of guanine moieties in DNA. Simple repetitive probes, cosmids, PACs, and chromosome-specific painting probes were labeled by ULS and used in a series of multicolor fluorescence in situ hybridization experiments on interphase and metaphase cells. It is demonstrated that ULS-labeled probes, in general, perform as well as the more conventional enzymatically labeled probes. The advantage of ULS labeling over enzymatic labeling techniques is that it is a fast and simple procedure, and that the labeling can easily be scaled up for bulk probe synthesis. In addition, with ULS labeling it is possible to label degraded DNA, a situation in which enzymatic labeling is known to perform unsatisfactorily.
Wiegant
J C
JC
Laboratory for Cytochemistry and Cytometry, Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. J.C.A.G.Wiegant@mcb.Medfac.LeidenUniv.NL
van Gijlswijk
R P
RP
Heetebrij
R J
RJ
Bezrookove
V
V
Raap
A K
AK
Tanke
H J
HJ
D001402:B-Lymphocytes / Q000473:pathology; D002503:Centromere / Q000235:genetics; D020223:Chromosome Painting / Q000295:instrumentation / Q000379:methods; D002877:Chromosomes, Human / Q000235:genetics; D002945:Cisplatin / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D003360:Cosmids / Q000235:genetics; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D006147:Guanine / Q000378:metabolism*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation / Q000379:methods*; D015463:Leukemia, Prolymphocytic / Q000235:genetics / Q000473:pathology; D008297:Male; D009030:Mosaicism / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014178:Translocation, Genetic / Q000235:genetics; D014407:Tumor Cells, Cultured
D003432:Cross-Linking Reagents; D015342:DNA Probes; D005456:Fluorescent Dyes; D006147:Guanine; D004247:DNA; D002945:Cisplatin