Human chromosome 5 sequence primer amplifies Alu polymorphisms on chromosomes 2 and 17.
Members of the Alu family of repetitive elements occur frequently in the human genome and are often polymorphic. Techniques involving Alu element mediated polymerase chain reactions (Alu PCR) allow the isolation of region-specific human DNA fragments from mixed DNA sources. Such fragments are a source of region-specific Alu elements useful for the detection of Alu-related polymorphisms. A clone from human chromosome 5, corresponding to locus D5F40S1, was isolated using Alu PCR differential hybridization. Alu elements within this clone were investigated for the presence of potentially polymorphic 3' polyA tails. Primers were devised to amplify the 3' polyA tail of an Alu element present within the clone. One primer, D5F40S1-T, was specific to the DNA flanking the 3' end of the Alu element, and the other primer was homologous to sequences within the element. When these primers were used in PCR reactions, products from chromosomes 2 and 17 (loci D2F40S2 and D17F40S3) were amplified in addition to the expected product from chromosome 5. The most likely explanation for this nonspecific amplification is that the D5F40S1-T primer is located within a low-copy repetitive element that is 3' of the Alu element. This phenomenon presents a potential problem for the identification of region-specific Alu polymorphisms.
Bernard
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Department of Medical Genetics, University of British Columbia, Vancouver, Canada.
Wood
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D000818:Animals; D001483:Base Sequence; D002886:Chromosomes, Human, Pair 17; D002895:Chromosomes, Human, Pair 5; D002897:Chromosomes, Human, Pair 7; D006224:Cricetinae; D003412:Cricetulus; D004277:DNA, Single-Stranded; D005260:Female; D008040:Genetic Linkage; D006801:Humans; D006822:Hybrid Cells; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D011061:Poly A; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid
D004277:DNA, Single-Stranded; D011061:Poly A