Effect of internal direct and inverted Alu repeat sequences on PCR.
We have studied the effect of repeated DNA sequence, especially Alu repeats, on PCR. Alu repeats are sequences that are approximately 300 bp long and interspersed at a very high copy number throughout the human genome. We amplified part of the human low-density lipoprotein receptor gene containing two Alu repeat sequences in the same orientation, approximately 7.8 kb apart, with unique sequence primers outside these repeats. The major PCR product was a DNA fragment with an in vitro deletion between the Alu repeats. The formation of this product depended on the template concentration and the type of polymerase used. Such a product arose apparently as a result of a "jumping reaction" involving a primer whose extension was terminated prematurely within one Alu repeated followed by annealing of such an incompletely extended primer to the other, distant Alu repeat. No such jumping products were seen when a 0.8-kb region containing two nearby inverted Alu repeats within the human alpha-galactosidase A gene was subject to PCR with unique sequence primers annealing just outside these repeats.
Ji
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Department of Biochemistry, Tulane Medical School, New Orleans, Louisiana 70112, USA.
Zhang
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Warshamana
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Qu
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Ehrlich
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D000818:Animals; D016477:Artifacts; D001483:Base Sequence; D004247:DNA; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats; D011973:Receptors, LDL / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D017384:Sequence Deletion
D017931:DNA Primers; D011973:Receptors, LDL; D004247:DNA