Prenatal screening
OwnerLawrence J. Wanghinventor
Life of this patent
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Abstract
The present invention concerns products and methods particularly useful for activating and analyzing non-dividing cell nuclei. The featured products include activating egg extracts, cytostatic factor (CSF) extracts, kits containing these extracts, and a microchamber microscope slide useful in analyzing nucleus activation.
Claims86
A method for activating a nucleus from a human fetal cell comprising the steps of:
- a) separating said nucleus from its surrounding cytoskeleton to form a pretreated nucleus, and
- b) contacting said pretreated nucleus with an activating egg extract to activate said pretreated nucleus.
A method of causing a non-dividing human nucleus to activate, using activating egg extract and CSF extract prepared from hardened eggs comprising:
- a) incubating said non-dividing human nucleus with said CSF extract prepared from hardened eggs to form a pretreated nucleus, and
- b) contacting said pretreated nucleus with said activating egg extract wherein said activating egg extract is prepared from synchronously activated hardened eggs.
A method for preparing an activating egg extract, comprising the steps of hardening a plurality of eggs, simultaneously inducing said eggs;
- and preparing an activating egg extract from said eggs wherein said eggs are induced for a length of time such that they have at least 70% of maximal activation DNA synthesis activity.
A method of causing a non-dividing cell nucleus to swell comprising the steps of:
- a) separating said nucleus from its surrounding cytoskeleton to form a pretreated nucleus;
- and
- b) contacting said pretreated nucleus with a CSF extract supplemented with an aqueous solution or a protein kinase inhibitor.
A method of causing chromosome formation in a non-dividing cell nucleus comprising the steps of:
- a) separating said nucleus from its surrounding cytoskeleton to form a pretreated nucleus;
- and
- b) contacting said pretreated nucleus with a CSF extract supplemented with a cyclin.
A method for activating or studying a mammalian sperm cell nucleus comprising the steps of:
- a) pretreating said sperm cell nucleus to form a pretreated sperm nucleus wherein said pretreating comprises (i) separating said nucleus from its surrounding cytoskeleton by permeabilizing said cell nuclear membrane and incubating in the presence of a protease and (ii) incubating in the presence of a thiol reducing agent;
- and
- b) activating said pretreated sperm cell.
The method of clam 34, wherein said human sperm cell nucleus is obtained from either a person infected with a virus or a person not infected with said virus.
An activation assay for studying male fertility comprising:
- a) pretreating a sperm nucleus to separate cytoskeletal protein from nucleic acid,
- b) activating said sperm nucleus,
- c) measuring activation of said nucleus activated in step (b).
A viral integration assay comprising the steps of:
- a) pretreating a cell nucleus to separate the nucleus from its surrounding cytoskeleton to form a pretreated nucleus,
- b) activating said nucleus and incubating with a viral integration complex containing viral nucleic acid, wherein said integration complex is added before or after said incubating;
- and
- c) measuring integration of viral nucleic acid into nucleic acid of said cell nucleus.
A viral integration assay comprising the steps of:
- a) constructing a pseudonucleus from a defined DNA template,
- b) activating said nucleus and incubating with a viral integration complex containing viral nucleic acid, wherein said integration complex is added before or after said incubating;
- and
- c) measuring integration of viral nucleic acid into nucleic acid of said cell nucleus.
A product for preparing a non-dividing nucleus to activate upon subsequent treatment with activating egg extract, comprising a CSF extract prepared using an eukaryotic cell, wherein said CSF extract is supplemented with Ca2+.
A product for causing nuclear swelling comprising CSF extract supplemented with a protein kinase inhibitor and/or an aqueous solution.
A product for causing chromosome formation in a cell nucleus comprising a CSF extract supplemented with a cyclin.
A product for activating a non-dividing nucleus comprising an activating egg extract having at least 70% optimal activation activity.
A kit for activating a non-dividing nucleus comprising a first product comprising frozen activating egg extract having at least 70% optimal activation activity and a second product comprising a frozen CSF extract.
A microscope slide comprising;
- a) an upper surface,
- b) a water repellent-means having a defined thickness located upon said upper surface to define a microchamber connected by a channel to at least one well on said upper surface, and
- c) said microchamber shaped to enhance flushing of said microchamber.
Dependencies are inferred from the claim text.