Genotyping the baboon ABO histo-blood group locus by two-color fluorescence SSCP.
The use of baboons in experimental medicine, especially as organ and tissue donors, would be facilitated by the availability of ABO histo-blood group O animals, which are currently rare. To meet our need in breeding and identifying such animals, we have developed a single-stranded conformational polymorphism (SSCP)-based genotyping assay using fluorescence detection. Various buffers and labeling schemes were evaluated to optimize allele discrimination. Through the use of a nested PCR protocol, single-cell sensitivity was achieved, making the assay applicable to preimplantation diagnosis following in vitro fertilization. We discuss the comparative advantages of SSCP vs. alternative methodologies for genotyping.
Diamond
D C
DC
Center for Molecular Biology and Gene Therapy, Loma Linda University and Medical Center, CA 92354, USA. rddiamond@earthlink.net
Illes
K
K
Bailey
L L
LL
Szalay
A A
AA
D000017:ABO Blood-Group System / Q000235:genetics*; D000483:Alleles; D000818:Animals; D002021:Buffers; D002478:Cells, Cultured; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D005838:Genotype; D010215:Papio; D016133:Polymerase Chain Reaction / Q000379:methods; D018807:Polymorphism, Single-Stranded Conformational
D000017:ABO Blood-Group System; D002021:Buffers; D017931:DNA Primers