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US 7,604,998 B2Granted
Fingerprint analysis for a plurality of oligonucleotides
AssigneeIntegrated Dna Technologies, Inc.
InventorBrian Elliott
Life of this patent
Estimating the term…
Abstract
The invention provides a method for evaluating the accuracy of an oligonucleotide sample, specifically a sample containing a variety of oligonucleotides of potentially varying size and sequence. The method provides a fingerprint that can be used to evaluate the accuracy of a multi-oligonucleotide sample whether or not the sample contains differing oligonucleotides that have the same or about the same molecular weight.
Claims11
A method of detecting the presence or absence of a constituent oligonucleotide in an oligonucleotide mixture, the method comprising:
- a) obtaining a measurement of a mass spectrum of the oligonucleotide mixture;
- b) comparing the measurement of the mass spectrum having a plurality of measured peaks with a calculated mass spectrum having a plurality of calculated peaks, said calculated mass spectrum derived from Formula 1:
- MWnew=(MW1*Intensity1+MW2*Intensity2+ . . . MWn*Intensityn)/(Intensity1+Intensity2+ . . . Intensityn)
- wherein MWnew is a calculated molecular weight of a calculated peak, MW1 is a first molecular weight of a first subset of one or more oligonucleotides in the oligonucleotide mixture, MW2 is a second molecular weight of a second subset of one or more oligonucleotides in the oligonucleotide mixture, MWn represents one or more additional molecular weights of one or more additional n subsets in the oligonucleotide mixture, Intensity1 is a first peak intensity of the first subset of one or more oligonucleotides in the oligonucleotide mixture, Intensity2 is a second peak intensity of the second subset of one or more oligonucleotides in the oligonucleotide mixture and Intensityn represents one or more additional peak intensities of the one or more additional n subsets in the oligonucleotide mixture;
- and
- c) detecting the presence or absence of the constituent oligonucleotide by correlating one or more of the measured peaks with one or more of the calculated peaks.
3 more elements
A method for predicting a measured peak intensity and a measured peak mass in an electrospray ionization mass spectroscopy trace, the method comprising:
- a) assigning each oligonucleotide member in a sample an initial intensity value of one;
- b) determining from a set of oligonucleotide members a first sub-set of oligonucleotide members that have an equal molecular weight;
- c) combining members of the first sub-set to form a combined first peak intensity using a formula:
- Inew=I1+I2+ . . . In
- wherein Inew is the first combined peak intensity and I1 and I2 are intensities of the oligonucleotide members of the first sub-set and In represents additional oligonucleotide members that may or may not be present in the first sub-set that have an equal molecular weight as I1 and I2;
- d) assigning a first combined peak mass to the first combined peak intensity that correlates with the molecular weight of each member of the first sub-set;
- e) determining from the set of oligonucleotide members a second sub-set of oligonucleotide members, said second sub-set of oligonucleotide members having a molecular weight within a range of other members of the second sub-set that when each member of the second sub-set is plotted as a peak in an electrospray ionization mass spectroscopy trace the peaks of members of the second sub-set overlap at greater than 50 percent peak height;
- f) combining members of the second sub-set to form a second combined peak intensity using a formula:
- I2new=I21+I22+ . . . I2n
- wherein I2new is the second combined peak intensity and I21 and I22 are intensities of the oligonucleotide members of the second sub-set and I2n represents additional oligonucleotide members that may or may not be present in the second sub-set;
- g) combining the members of the second sub-set to form a second combined peak mass using the formula:
- MWnew=(MW1*I21+MW2*I22+ . . . MWn*I2n)/(I21+I22+ . . . I2n)
- wherein MWnew represents the second combined peak mass, MW1 and MW2 are the molecular weights of members of the second sub-set, and MWn represents additional n members that may be present in the second sub-set;
- and
- predicting a measured peak intensity and a measured peak mass in an electrospray ionization mass spectrometry trace from the second combined peak intensity and the second combined peak mass.
12 more elements
Dependencies are inferred from the claim text.