Preparation of nucleic acid samples
First Claim
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1. A method of preparing labeled fragments of a population of nucleic acids of interest comprising:
- increasing the relative percentage of said population of nucleic acids of interest within a mixed population of nucleic acids, wherein said mixed population comprises a plurality of nucleic acid sequences, comprising;
(a) contacting a nucleic acid sample with a nucleic acid bait molecule, wherein said nucleic acid sample comprises said nucleic acids of interest and at least one unwanted target sequence and wherein said bait molecule binds specifically to said unwanted target sequence, but not to said nucleic acids of interest, under such conditions as to allow for the formation of a bait;
target complex;
(b) digesting the unwanted target sequence in the bait;
target complex thereby resulting in an increase in the relative percentage of said nucleic acids of interest within said mixed population of nucleic acids;
fragmenting said nucleic acids of interest to produce fragments; and
adding a label to the fragments.
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Abstract
The presently claimed invention provides methods, compositions, and apparatus for studying nucleic acids. Specifically, the present invention provides a novel enrichment and labeling strategy for ribonucleic acids. In one embodiment, the invention provides enriching for a population of interest in a complex population by diminishing the presence of a target sequence. In a further embodiment, the invention can be used to reproducibly label and detect extremely small amounts of nucleic acids.
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Citations
41 Claims
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1. A method of preparing labeled fragments of a population of nucleic acids of interest comprising:
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increasing the relative percentage of said population of nucleic acids of interest within a mixed population of nucleic acids, wherein said mixed population comprises a plurality of nucleic acid sequences, comprising;
(a) contacting a nucleic acid sample with a nucleic acid bait molecule, wherein said nucleic acid sample comprises said nucleic acids of interest and at least one unwanted target sequence and wherein said bait molecule binds specifically to said unwanted target sequence, but not to said nucleic acids of interest, under such conditions as to allow for the formation of a bait;
target complex;
(b) digesting the unwanted target sequence in the bait;
target complex thereby resulting in an increase in the relative percentage of said nucleic acids of interest within said mixed population of nucleic acids;
fragmenting said nucleic acids of interest to produce fragments; and
adding a label to the fragments. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, 41)
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33. The method of 32 wherein a thermostable RNAse H is used to remove said target sequences from said bait:
- target complex.
Specification