Compositions and methods of use of standardized mixtures
AssigneesMedical College of Ohio, Gene Express, Inc.
InventorsJames Willey, Brad Austermiller, Erin Crawford, Charles Knight, Terry Osborn, Robert Zahorchak
Life of this patent
Estimating the term…
Abstract
The present invention is directed to methods and compositions for evaluating nucleic acids, methods of preparing such compositions, and applications and business methods employing such compositions and methods. In particular, the present invention provides business methods for operating a gene expression measurement service.
Claims40
A method of assessing a first nucleic acid in a first sample, comprising:
- providing a standardized mixture comprising a competitive template for said first nucleic acid and a competitive template for a second nucleic acid in said first sample wherein said competitive templates are at known concentrations relative to each other;
- combining said first sample with said standardized mixture;
- co-amplifying said first nucleic acid and said competitive template for said first nucleic acid to produce first amplified product thereof;
- diluting said first amplified product;
- further co-amplifying said diluted first amplified product of said first nucleic acid and of said competitive template for said first nucleic acid, to produce second amplified product thereof;
- and
- co-amplifying said second nucleic acid and said competitive template for said second nucleic acid to produce first amplified product thereof.
A method for assessing a first nucleic acid, comprising:
- providing a series of serially-diluted standardized mixtures comprising a competitive template for said first nucleic acid and a competitive template for a second nucleic acid present in a number of samples comprising said first nucleic acid, wherein said competitive templates are at known concentrations relative to each other;
- combining one of said samples comprising said first nucleic acid with a first one of said serially-diluted standardized mixtures;
- co-amplifying said first nucleic acid and said competitive template for said first nucleic acid to produce amplified product thereof;
- obtaining a first relationship, said first relationship comparing said amplified product of said first nucleic acid to said amplified product of said competitive template for said first nucleic acid;
- determining whether said first relationship is within about 1:10 to about 10:1;
- if not, repeating said combining, co-amplifying, obtaining and determining steps with a second one of said serially-diluted standardized mixtures;
- co-amplifying said second nucleic acid and said competitive template for said second nucleic acid to produce amplified product thereof;
- obtaining a second relationship, said second relationship comparing said amplified product of said second nucleic acid to said amplified product of said competitive template for said second nucleic acid;
- and
- comparing said first and said second relationships.
A method comprising preparing a standardized mixture of reagents, said reagents comprising sufficient competitive template for assessing amounts of a number of nucleic acids in more than about 106 samples wherein said standardized mixture allows direct comparison of said amounts between 2 of said samples.
A method comprising preparing a series of serially-diluted standardized mixtures of reagents, said reagent comprising sufficient competitive template for assessing amounts of a number of nucleic acids in more than about 106 samples wherein said standardized mixtures allow direct comparison of said amounts between 2 of said samples.
A composition comprising a standardized mixture of reagents, said reagents comprising sufficient competitive template for assessing amounts of a number of nucleic acids in more than about 106 samples wherein said standardized mixture allows direct comparison of said amounts between 2 of said samples.
A composition comprising a series of serially-diluted standardized mixtures of reagents, said reagent comprising sufficient competitive template for assessing amounts of a number of nucleic acids in more than about 106 samples wherein said standardized mixtures allow direct comparison of said amounts between 2 of said samples.
Dependencies are inferred from the claim text.