Chapter 32
Centromere-Specific Multicolor-FISH (cenM-FISH)
ANGELA NIETZEL, ANITA HELLER, HElKE STARKE, UWE CLAUSEN,
and THOMAS LIEHR
Introduction
24-color FISH using five different fluorochromes has recently been described as M-FISH or SKY, and is a useful technique for the characterization of complex chromosomal aberrations (Speicher et aI., 1996; Schrock
et aI., 1996; for an overview of multicolor-FISH literature see http://mtin.mtLuni-jena.de!-huwww!MoLZYTO!mFISHlit.htm). For technical reasons whole or partial chromosome libraries, each specific for an entire
chromosome, exclude the centromeric regions of the color staining due
to the blocking of repetitive sequences by COTl-DNA. Here, we describe
a recently developed new multicolor FISH technique - called centromerespecific multicolor-FISH (cenM-FISH) (Nietzel et a1. 2001). The technique
is based on all available centromere-specific DNA probes, which are labeled in five different fluorochromes (i.e. TexasRed, SpectrumGreen,
SpectrumOrange, CyS and DEAC) according to Fig. IB and hybridized
simultaneously. This approach allows the identification of all human centromeres by their individual coloring in one single step and therefore is a
powerful tool in molecular cytogenetics. Centromere-specific multicolor
FISH is a very helpful technique especially for the characterization of small
Angela Nietzel, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
Anita Heller, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
Heike Starke, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
Uwe Clausen, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
~ Thomas Liehr, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany (phone +49-3641-935533; fax +49-3641-935502;
e-mail i8Iith@mti-n.mtLuni-jena.de)
PROTOCOL
Centromere-Specific Multicolor-FISH (cenM-FISH)
ANGELA NIETZEL, ANITA HELLER, HElKE STARKE, UWE CLAUSEN,
and THOMAS LIEHR
Introduction
24-color FISH using five different fluorochromes has recently been described as M-FISH or SKY, and is a useful technique for the characterization of complex chromosomal aberrations (Speicher et aI., 1996; Schrock
et aI., 1996; for an overview of multicolor-FISH literature see http://mtin.mtLuni-jena.de!-huwww!MoLZYTO!mFISHlit.htm). For technical reasons whole or partial chromosome libraries, each specific for an entire
chromosome, exclude the centromeric regions of the color staining due
to the blocking of repetitive sequences by COTl-DNA. Here, we describe
a recently developed new multicolor FISH technique - called centromerespecific multicolor-FISH (cenM-FISH) (Nietzel et a1. 2001). The technique
is based on all available centromere-specific DNA probes, which are labeled in five different fluorochromes (i.e. TexasRed, SpectrumGreen,
SpectrumOrange, CyS and DEAC) according to Fig. IB and hybridized
simultaneously. This approach allows the identification of all human centromeres by their individual coloring in one single step and therefore is a
powerful tool in molecular cytogenetics. Centromere-specific multicolor
FISH is a very helpful technique especially for the characterization of small
Angela Nietzel, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
Anita Heller, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
Heike Starke, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
Uwe Clausen, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany
~ Thomas Liehr, Institute of Human Genetics and Anthropology, Kollegiengasse 10,
Jena, 07743, Germany (phone +49-3641-935533; fax +49-3641-935502;
e-mail i8Iith@mti-n.mtLuni-jena.de)
PROTOCOL