Detection of universal variable fragments as markers for genetic studies. A novel technology for DNA fingerprinting.
A novel DNA technology enables the detection of universal variable fragments (UVF), thus revealing genetic variation without a priori sequence information. The detection of UVF markers is based on two amplifications of genomic DNA with the polymerase chain reaction. In the first amplification, two short oligonucleotide primers produce a large number of fragments. One primer is based on a microsatellite sequence, whereas the second primer can have any sequence. In the second amplification, the length of the primers is increased in order to decrease the number of amplicons. This enables the selection of polymorphic fragments. Restriction digestion can be used to further increase the number of polymorphisms. Until now, we have demonstrated UVF in several different species. In addition, with the present study we have contributed to the linkage map of the rabbit by localizing 11 UVF markers on different linkage groups. Mendelian inheritance was shown in this linkage study through a backcross of two inbred rabbit strains. The power of the UVF technique is based on the selection for microsatellite variation in combination with the detection of single-nucleotide polymorphisms. UVF thus offers the possibility of increasing the clustering of markers and localizing genes in species for which sequence information is either not present or only scarcely present.
van Haeringen
W A
WA
University of Utrecht, Department of Laboratory Animal Science, PO Box 80166, 3508 TD, Utrecht, The Netherlands. info@vhlgenetics.com
van de Goor
L H P
LH
Panneman
H
H
van Lith
H A
HA
van Haeringen
H
H
van Zutphen
L F M
LF
D000818:Animals; D017849:Animals, Inbred Strains; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D005240:Feasibility Studies; D020869:Gene Expression Profiling / Q000379:methods*; D008040:Genetic Linkage; D005819:Genetic Markers / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics; D006736:Horses / Q000235:genetics; D018895:Microsatellite Repeats / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics; D011817:Rabbits / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D017931:DNA Primers; D005819:Genetic Markers