Painting of human chromosomes with probes generated from hybrid cell lines by PCR with Alu and L1 primers

Specific amplification of human sequences of up to several kb
length has recently been accomplished in man-hamster and man-mouse
somatic hybrid cell DNA by IRS-PCR (interspersed repetitive
sequence — polymerase chain reaction). This approach is based on
oligonucleotide primers that anneal specifically to human Alu- or
L1-sequences and allows the amplification of any human sequences
located between adequately spaced, inverted Alu- or L1-blocks.
Here, we demonstrate that probe pools generated from two somatic
hybrid cell lines by Alu- and L1-PCR can be used for chromosome
painting in normal human lymphocyte metaphase spreads by
chromosomal in situ suppression (CISS-) hybridization. The painted
chromosomes and chromosome subregions directly represent the
content of normal and deleted human chromosomes in the two somatic
hybrid cell lines. The combination of IRS-PCR and
CISS-hybridization will facilitate and improve the cytogenetic
analysis of somatic hybrid cell panels, in particular, in cases
where structurally aberrant human chromosomes or human chromosome
segments involved in interspecies translocations cannot be
unequivocally identified by classical banding techniques. Moreover,
this new approach will help to generate probe pools for the
specific delineation of human chromosome subregions for use in
cytogenetic diagnostics and research without the necessity of
cloning.