A PCR-based method to amplify DNA with random primers: determining the chromosomal content of porcine flow-karyotype peaks by chromosome painting.
We present here a new PCR-based technique that allows the production of several micrograms of DNA from only 300 flow-sorted chromosomes. During the first two PCR cycles, the annealing temperature is decreased to 30 degrees C, and numerous random loci are amplified under nonspecific conditions. As demonstrated here for pig chromosomes 1 and 18, the PCR products may be used to identify the chromosomal content of the flow-karyotype peaks of any species by fluorescence in situ hybridization.
Milan
D
D
Laboratoire de Génétique Cellulaire, INRA, Castanet-Tolosan, France.
Yerle
M
M
Schmitz
A
A
Chaput
B
B
Vaiman
M
M
Frelat
G
G
Gellin
J
J
D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D004247:DNA / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000032:analysis; D005434:Flow Cytometry; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping / Q000379:methods*; D008214:Lymphocytes; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D013552:Swine / Q000235:genetics*
D004277:DNA, Single-Stranded; D004247:DNA