Polymerase Chain reaction generated probes for fluorescence in situ hybridization.
The extended use of Fish with centromeric probes in many cytogenetic laboratories is often impaired by the cost of this technique. Polymerase Chain Reaction (PCR) constitutes a simple way to generate and label such centromeric probes at low cost. Two types of human DNA source can be used: 1--Somatic hybrid cell lines containing a unique human chromosome. The specific amplification of the human subset of alphoid DNA is realised with a primer pair specific for the consensus region of human alpha satellite sequence. 2--Total Human DNA. This time, a primer pair specific for the alpha satellite DNA of the chromosome of interest must be designed. These probes, labelled during the PCR reaction by direct incorporation of modified dUTP, are actually widely used in our laboratory, alone or mixed with other probes (chromosome painting or locus specific probes).
Dupont
J M
JM
Laboratoire d'Histologie Embryologie Cytogénétique, GH Cochin, 123 Bd Port-Royal, 75014 Paris.
Lebbar
A
A
Dupuy
O
O
Frydman
N
N
Letessier
D
D
Auvinet
P
P
Rabineau
D
D
D002478:Cells, Cultured; D015342:DNA Probes; D006801:Humans; D006822:Hybrid Cells; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
D015342:DNA Probes