Appeal No. 2001-1880
Application No. 09/273,835
(c) measuring the mass peak of the labeled oligonucleotide using mass
spectrometry;
(d) obtaining the magnitude of the mass shift between the labeled
oligonucleotide and the unlabeled oligonucleotide, whereby the number of isotope-
labeled nucleotides in the oligonucleotide under investigation is determined; and
(e) comparing the number of isotope-labeled nucleotides with the number of that
type of nucleotide in a reference oligonucleotide.
4. The method for determining the nucleotide composition of an oligonucleotide
as described in claim 3, wherein the chosen primers contain a sequence for the type IIS
restriction enzyme.
5. The method for determining the nucleotide composition of an oligonucleotide
as described in claim 1, wherein said step of incorporating a stable, isotope-labeled
form of one of the four nucleotide units of an oligonucleotide into the oligonucleotide
under investigation in place of the ordinary nucleotide therein is achieved using
isothermal rolling-circle amplification.
8. A method for determining the nucleotide composition of an oligonucleotide
which comprises the steps of:
(a) incorporating a stable, isotope-labeled form of two of the four nucleotide units
of an oligonucleotide into the oligonucleotide under investigation in place of the ordinary
nucleotide therein, the other three types of nucleotides in the oligonucleotide being
unlabeled;
(b) measuring the mass peak of the unlabeled oligonucleotide using mass
spectrometry;
(c) measuring the mass peak of the labeled oligonucleotide using mass
spectrometry;
(d) obtaining the magnitude of the mass shift between the labeled
oligonucleotide and the unlabeled oligonucleotide, whereby the number of isotope-
labeled nucleotides in the oligonucleotide under investigation is determined; and
(e) comparing the number of isotope-labeled nucleotides with the number of that
type of nucleotide in a reference oligonucleotide.
14. A method for detecting polymorphisms in oligonucleotides which comprises
the steps of:
(a) incorporating a stable, isotope-labeled form of one of the four nucleotide units
of an oligonucleotide into the oligonucleotide under investigation in place of the ordinary
nucleotide therein, the other three types of nucleotides in the oligonucleotide being
unlabeled;
(b) measuring the mass peak of the unlabeled oligonucleotide using mass
spectrometry;
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