Methods and kits for isolating cells
AssigneePromega Corporation
InventorsRyan J. Olson, Rex M. Bitner, Allan M. Tereba, Laura L. Bozek
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Abstract
Disclosed are methods for differential extraction of a target component from a sample which is predominantly composed of other types of non-target cells that can be lysed using methods that do not lyse the target cells, so that the target material can be purified away from the lysed non-target material. One exemplary method is directed to isolating sperm cells from an aqueous sample and kits for performing same.
Claims44
A method of isolating first cells from a sample comprising first cells and second cells comprising:
- (a) treating the sample to form an aqueous lysate under conditions that lyse the second cells without lysing the first cells;
- (b) applying a force to the sample for a period of time sufficient to form a pellet comprising the first cells;
- and
- (c) forming a pellet immobilizing cap between the pellet and the aqueous lysate.
A method of isolating sperm cell DNA from an aqueous sample comprising epithelial cells comprising:
- (a) treating the sample under conditions that allow selective lysis of the epithelial cells;
- (b) applying a force to the treated sample for a period of time sufficient to form a sperm pellet;
- (c) forming a pellet immobilizing cap between the sperm pellet and the aqueous material containing lysed epithelial cells;
- (d) removing the aqueous material;
- and
- (e) lysing the cells in the sperm pellet under conditions that allow sperm cell DNA purification.
A method of separating intact cells or target organelles from an aqueous cell lysate sample comprising:
- (a) applying a force to the sample for a period of time sufficient to form a pellet comprising at least some of the intact cells or target organelles;
- and
- (b) forming a pellet immobilizing cap between the pellet and the aqueous cell lysate.
A method of isolating target organelles from a sample comprising cells comprising a target organelle comprising:
- (a) treating the sample to form an aqueous lysate under conditions that selectively lyse the cells without lysing the organelles;
- (b) applying a force to the sample for a period of time sufficient to form a pellet comprising the target organelles;
- and
- (c) forming a pellet immobilizing cap between the pellet and the lysate.
A method of isolating a target cell substituent from a sample comprising target cells comprising:
- (a) treating the sample to form an aqueous lysate under conditions that selectively lyse the target cells and under which at least one organelle is not lysed;
- (b) applying a force to the sample for a period of time sufficient to form a pellet comprising the unlysed organelles;
- and
- (c) forming a pellet immobilizing cap between the pellet and the lysate.
A kit for isolating a target cell or organelle from a non-target cell comprising a lysis agent for preferentially lysing the non-target cell and a pellet immobilizing material.
Dependencies are inferred from the claim text.