Gel microdroplets in genetic analysis
First Claim
1. A method of nucleic acid analysis, comprising forming a population of gel microdrops encapsulating a population of biological entities, each entity comprising a nucleic acid, whereby at least some microdrops in the population each encapsulate a single entity;
- contacting the population of gel microdrops with a probe under conditions whereby the probe specifically hybridizes to at least one complementary sequence in the nucleic acid in at least one gel microdrop;
isolating or detecting the at least one gel microdrop.
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Abstract
The invention provides methods of nucleic acid analysis. Such methods entail forming a population of gel microdrops encapsulating a population of biological entities, each entity comprising a nucleic acid, whereby at least some microdrops in the population each encapsulate a single entity. The population of gel microdrops is then contacted with a probe under conditions whereby the probe specifically hybridizes to at least one complementary sequence in the nucleic acid in at least one gel microdrop. At least one gel microdrop is then analyzed or detected. The biological entities can be cells, viruses, nuclei and chromosomes.
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Citations
68 Claims
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1. A method of nucleic acid analysis, comprising forming a population of gel microdrops encapsulating a population of biological entities, each entity comprising a nucleic acid, whereby at least some microdrops in the population each encapsulate a single entity;
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contacting the population of gel microdrops with a probe under conditions whereby the probe specifically hybridizes to at least one complementary sequence in the nucleic acid in at least one gel microdrop;
isolating or detecting the at least one gel microdrop. - 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, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49)
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50. A method of diagnosing a disease due to a mutation, comprising:
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obtaining a sample of cells from a patient;
encapsulating a population of chromosomes from the sample in a population of microdrops;
contacting the microdrops with a first probe that is complementary to a nucleic acid segment containing the somatic mutation, and a second probe complementary to the chromosome in which the somatic mutation occurs at a site distal to the somatic mutation, whereby the first probe hybridizes to microdrops bearing the chromosome with a somatic mutation and the second probe hybridizes to microdrops bearing the chromosome irrespective whether the somatic mutation is present;
determining the ratio of microdrops hybridizing to the first probe and hybridizing to the second probe;
diagnosing the existence or prognosis of the disease from the ratio. - View Dependent Claims (51, 52)
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53. A method of chromosome analysis, comprising forming a population of gel micropdrops encapsulating a population of nucleic, whereby at least some microdrops in the population each encapsulate a single nucleus;
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contacting the population of gel microdrops with a probe under conditions whereby the probe specifically hybridizes to at least one complementary sequence in at least one chromosome in a nucleus of least one gel microdroplet;
isolating or detecting the at least one gel microdroplet.
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54. A method of isolating chromosomes comprising:
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culturing a population of cells in genistein and colcemid to synchronize chromosomes in metaphase;
isolating chromosomes from the cells.
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55. A method of isolating chromosomes comprising:
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lysing a population of cells to form a lysate;
treating the lysate with an antibody linked to a magnetic particles, wherein the antibody specifically binds to one or more chromosomes in the cells;
isolating magnetic particles from the lysate.
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56. A method of chromosome analysis, comprising forming a population of gel micropdrops encapsulating a population of cells or nuclei, whereby at least some microdrops in the population each encapsulate a single nucleus;
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contacting the population of gel microdrops with a probe under conditions whereby the probe specifically hybridizes to at least one complementary sequence in at least one nucleus in at least one gel microdrop;
isolating or detecting the at least one gel microdrop. - View Dependent Claims (57, 58, 59, 60, 61, 62, 63, 64, 65, 66)
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- 67. A kit comprising a high melting temperature agarose, emulsification equipment, a label indicating how to use the kit for probe hybridization analysis.
Specification