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US 10,370,649 B2Granted
Device and method for cell nuclei preparation
AssigneeGe Healthcare UK Limited
InventorsMichael Kenneth Kenrick, Geraint Seymour
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Abstract
The present invention relates to the field of nucleic acid collection and storage, and in particular to the collection and long-term storage of nuclei. The invention provides a device and method which can be used to capture and store nuclei at ambient temperatures allowing the subsequent isolation of nucleic acids by passive washing with a wash buffer. The present invention also provides a kit including the device of the invention suitable for carrying out the method of the invention.
Claims32
A device comprising:
- (i) a container comprising an open upper end, a sealed lower end and a chamber therebetween defined by chamber walls;
- wherein said container further comprises a hollow tube adjacent the upper end allowing gaseous and liquid communication with the chamber and a removable base sealing the lower end, and wherein said chamber contains a cell lysis reagent;
- (ii) a plunger comprising a hollow plunger body defined by plunger walls wherein said plunger walls have a shape conforming to and smaller than said chamber to allow a bottom end of the plunger to sealably slide within the chamber against the chamber walls, a septum cap sealing a top end of said plunger body, and a solid support spanning an interior transverse section of said plunger body, wherein said solid support is a permeable membrane;
- and
- (iii) a vacuum port connected to the hollow tube, wherein negative pressure is created within the chamber through the hollow tube attached to the vacuum port.
2 more elements
A method comprising:
- (i) introducing a cellular sample into a container wherein said container has an open upper end and sealed lower end and a chamber therebetween defined by chamber walls, wherein said container further comprises a hollow tube adjacent the upper end allowing gaseous and liquid communication with the chamber, wherein negative pressure is created within the chamber through a vacuum port attached to the hollow tube, wherein said container contains a collection buffer comprising a lysis reagent at a concentration sufficient to selectively lyse the cytoplasmic membranes and a small proportion of the nuclear membranes present in the cellular sample and leave a large proportion of the cell nuclei intact;
- (ii) inserting a plunger into the upper end of said container wherein said plunger comprises a hollow plunger body defined by plunger walls, wherein said walls have a shape conforming to and smaller than said chamber, a septum cap sealing the top end of said plunger body, and a solid support comprising a permeable membrane spanning an interior transverse section of said plunger body;
- (iii) pressing said plunger towards said lower end of said container so that said solid support passes through said cellular sample and the cell nuclei and other cellular components bind to the solid support leaving filtrate behind;
- and,
- (iv) removing said filtrate.
2 more elements
Dependencies are inferred from the claim text.