Faculty Research 1990 - 1999
Multilocus markers for mouse genome analysis: PCR amplification based on single primers of arbitrary nucleotide sequence.
Document Type
Article
Publication Date
1992
Keywords
Base-Composition, Base-Sequence, DNA, Genetic-Markers, Genome, Linkage-(Genetics), Magnesium, Mice, Mice-Inbred-Strains, Molecular-Sequence-Data, Polymerase-Chain-Reaction: mt, SUPPORT-NON-U-S-GOVT, SUPPORT-U-S-GOVT-NON-P-H-S, SUPPORT-U-S-GOVT-P-H-S, Templates, Variation-(Genetics)
First Page
55
Last Page
64
JAX Source
Mamm Genome 1992;3(2):55-64
Grant
HG00189, HL32087
Abstract
Polymerase chain reaction (PCR) based on single primers of arbitrary nucleotide sequence provides a powerful marker system for genome analysis because each primer amplifies multiple products, and cloning, sequencing, and hybridization are not required. We have evaluated this typing system for the mouse by identifying optimal PCR conditions; characterizing effects of GC content, primer length, and multiplexed primers; demonstrating considerable variation among a panel of inbred strains; and establishing linkage for several products. Mg2+, primer, template, and annealing conditions were identified that optimized the number and resolution of amplified products. Primers with 40% GC content failed to amplify products readily, primers with 50% GC content resulted in reasonable amplification, and primers with 60% GC content gave the largest number of well-resolved products. Longer primers did not necessarily amplify more products than shorter primers of the same proportional GC content. Multiplexed primers yielded more products than either primer alone and usually revealed novel variants. A strain survey showed that most strains could be readily distinguished with a modest number of primers. Finally, linkage for seven products was established on five chromosomes. These characteristics establish single primer PCR as a powerful method for mouse genome analysis.
Recommended Citation
Nadeau JH,
Bedigian HG,
Bouchard G,
Denial T,
Kosowsky M,
Norberg R,
Pugh S,
Sargeant E,
Turner R,
Paigen B.
Multilocus markers for mouse genome analysis: PCR amplification based on single primers of arbitrary nucleotide sequence. Mamm Genome 1992;3(2):55-64