Generation of mouse chromosome painting probes by DOP-PCR amplification of microdissected meiotic chromosomes.
We describe here the development of four sets of painting probes for mouse chromosomes 1 and 13, 2 and 8, 6 and 15, and X and Y by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) amplification of 10-20 copies of microdissected male meiotic chromosomes. The X,Y probe was obtained from the X-Y bivalent of diakinesis/metaphase I complements of mice with a normal karyotype, whereas the other probes were derived from tri- or quadrivalents in diakinesis/metaphase I of two reciprocal translocations, T(1;13)70H and T(2;8)2Wa, and one inversion heterozygote carrying a small deletion, In(6;15)Rb1Ald del6(15)1Wa. The specificity of these probes was established by fluorescence in situ hybridization (FISH) to meiotic and mitotic metaphase complements. The chromosomes painted by these probes could be identified by single or multicolor FISH.
Xiao
Y
Y
MGC-Department of Radiation Genetics and Chemical Mutagenesis, Leiden University, The Netherlands.
Darroudi
F
F
Kuipers
A G
AG
de Jong
J H
JH
de Boer
P
P
Natarajan
A T
AT
D000818:Animals; D002875:Chromosomes; D004396:Coloring Agents; D017931:DNA Primers; D015342:DNA Probes; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008540:Meiosis; D051379:Mice; D016133:Polymerase Chain Reaction; D013090:Spermatocytes; D014960:X Chromosome; D014998:Y Chromosome
D004396:Coloring Agents; D017931:DNA Primers; D015342:DNA Probes