Back to results
US 6,913,884 B2Granted
Compositions and methods for repetitive use of genomic DNA
AssigneeIllumina, Inc.
InventorsJohn R. Stuelpnagel, Mark S. Chee
Life of this patent
Estimating the term…
Abstract
The present invention relates to detection or genotyping (or other sample analysis) of target nucleic acids following immobilization of the target nucleic acids onto a surface. The target nucleic acids can be re-used multiple times, thus conserving sample materials and simplifying sample preparation.
Claims36
A method comprising:
- a) providing a composition camprising first primers and at least 10 different target nucleic acids, wherein either said first primers or said at least 10 different target nucleic acids are immobilized to at least one solid support;
- b) performing a first analysis of said at least 10 different target nucleic acids, said first analysis comprising:
- i) contacting said first primers with said at least 10 different target nucleic acids whereby at least one of said first primers hybridizes with said at least 10 different target nucleic acids;
- ii) removing unhybridized first primers;
- and
- iii) contacting said hybridized first primers with an enzyme such that said hybridized first primers are modified forming first modified primers, whereby a sinnal is obtained from said first analysis and whereby said at least 10 different target nucleic acids is not consumed, and
- c) performing a second analysis of said at least 10 different target nucleic acids, comprising:
- i) contacting second primers with said at least 10 different target nucleic acids whereby at least one of said second primers hybridizes with said at least 10 different target nucleic acids;
- ii) removing unhybridized second primers;
- and
- iii) contacting said hybridized second primers with an enzyme such that said hybridized second primers are modified forming second modified primers, whereby a signal is obtained from said second analysis and wherein a signal obtained from said second analysis is not diminished more than 40% compared to a signal obtained from said first analysis.
8 more elements
A method comprising:
- a) providing a composition comprising first primers and a target nucleic acid wherein said first primers are ligation primers;
- b) hybridizing said first ligation primers with said target nucleic acid to form first ligation complexes, whereby said first ligation primers hybridize to said target nucleic acid comprising a first target sequence;
- c) removing unhybridized ligation primers;
- d) contacting said first ligation complexes with a ligation enzyme, whereby when said first ligation primers are complementary to said first target sequence, said ligation enzyme ligates said first ligation primers generating first ligation products, whereby a signal is obtained from said first ligation products;
- e) removing said first ligation products from said target nucleic acid;
- f) hybridizing said target nucleic acid with second ligation primers to form second ligation complexes, whereby said second ligation primers hybridize to said target nucleic acid comprising a second target sequence, and
- g) contacting said second ligation complex with a ligation enzyme, whereby when said second ligation primers are complementary to said second target sequence, said ligation enzyme ligates said second ligation primers generating second ligation products, whereby a signal is obtained from said second ligation products and wherein a signal obtained from said second ligation products is not diminished more than 40% compared to a signal obtained from said first ligation products.
4 more elements
A method of reusing a target nucleic acid comprising:
- a) providing a composition comprising first primers and a target nucleic acid, wherein either said first primers or said target nucleic acid are immobilized on at least one solid support;
- b) performing a first analysis of said target nucleic acid, said first analysis comprising:
- i) contacting said first primers with said target nucleic acid whereby at least one of said first primers hybridizes with said target nucleic acid;
- ii) removing unhybridized first primers;
- and
- iii) contacting said hybridized first primers with an enzyme such that said hybridized first primers are modified forming first modified primers, whereby a signal is obtained from said first analysis and whereby said target nucleic acid is not consumed;
- and
- c) reusing said target nucleic acid in a second analysis, whereby a signal is obtained from said second analysis and wherein a signal obtained from said second analysis is not diminished more than 40% compared to a signal obtained from said first analysis.
5 more elements
Dependencies are inferred from the claim text.