Methods for measuring telomere length
First Claim
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1. A method for measuring an average length of telomeres in a sample comprising a cell or tissue, said method comprising steps of:
- (a) contacting said telomeres with an oligonucleotide linker under conditions such that said linker is covalently linked to a 3'"'"' end of said telomeres;
(b) amplifying with a first primer complementary to said oligonucleotide linker and a second primer complementary to a subtelomeric region of a chromosome under conditions in a reaction mixture such that said first and second primers hybridize specifically to said linker and subtelomeric region, respectively, and are extended by a polymerase, thereby forming first and second primer extension products; and
(c) measuring an amount by which said first and/or second primer has been extended to form said extension products or duplex nucleic acids comprising said first and second primer extension products to determine an average telomere length.
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Abstract
Methods are provided for the determination of telomere length. These methods can be used for diagnosis of cancer and the staging of cancer, and diagnosis of senecesence in cells. Also, the instant methods can be used to determine stages of diseases such as atherosclerosis or HIV infection, and can be used to diagnose infertility.
56 Citations
8 Claims
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1. A method for measuring an average length of telomeres in a sample comprising a cell or tissue, said method comprising steps of:
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(a) contacting said telomeres with an oligonucleotide linker under conditions such that said linker is covalently linked to a 3'"'"' end of said telomeres; (b) amplifying with a first primer complementary to said oligonucleotide linker and a second primer complementary to a subtelomeric region of a chromosome under conditions in a reaction mixture such that said first and second primers hybridize specifically to said linker and subtelomeric region, respectively, and are extended by a polymerase, thereby forming first and second primer extension products; and (c) measuring an amount by which said first and/or second primer has been extended to form said extension products or duplex nucleic acids comprising said first and second primer extension products to determine an average telomere length. - View Dependent Claims (3, 4, 5, 6, 7, 8)
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2. A method of diagnosing infertility in males, wherein said method comprises steps of:
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(a) measuring telomere length in a sperm sample from a subject to obtain a measured telomere length, by; (i) contacting telomeres in said sperm sample with an oligonucleotide linker under conditions such that said linker is covalently linked to a 3'"'"' end of said telomeres; (ii) amplifying with a first primer complementary to said oligonucleotide linker and a second primer complementary to a subtelomeric region of a chromosome under conditions such that said first and second primers hybridize specifically to said linker and subtelomeric region, respectively, and are extended by a polymerase, thereby forming first and second primer extension products; and (iii) measuring an amount by which said first and/or second primer has been extended to form said first and second primer extension products or duplex nucleic acids comprising said extension products to determine an average telomere length; (b) comparing said measured telomere length to a control telomere length obtained by measuring telomere length in a sperm sample obtained from a fertile male; and (c) correlating a decreased telomere length in said sample from said subject, as compared to said control telomere length, with infertility.
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Specification