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US 10,532,324 B1Granted
Instruments, modules, and methods for improved detection of edited sequences in live cells
AssigneeInscripta, Inc.
InventorsDon Masquelier, Phillip Belgrader, Andrew Garst, Richard Fox, Matthew Estes, Bruce Chabansky
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Abstract
The present disclosure provides instruments, modules and methods for improved detection of edited cells following nucleic acid-guided nuclease genome editing. The disclosure provides improved automated instruments that perform methods—including high throughput methods—for screening cells that have been subjected to editing and identifying cells that have been properly edited.
Claims20
A solid wall isolation, incubation and normalization (SWIIN) module comprising:
- a retentate member comprising;
- an upper surface and a lower surface and a first and second end,
- an upper portion of a serpentine channel defined by raised areas on the lower surface of the retentate member, wherein the upper portion of the serpentine channel traverses the lower surface of the retentate member for about 50% to about 90% of the length and width of the lower surface of the retentate member;
- at least one retentate port fluidically connected to the upper portion of the serpentine channel;
- and
- a reservoir cover at the first end of the retentate member;
- a permeate member disposed under the retentate member comprising:
- an upper surface and a lower surface and a first and second end,
- a lower portion of the serpentine channel defined by raised areas on the upper surface of the permeate member, wherein the lower portion of the serpentine channel traverses the upper surface of the permeate member for about 50% to about 90% of the length and width of the upper surface of the permeate member, and wherein the lower portion of the serpentine channel is configured to mate with the upper portion of the serpentine channel to form the serpentine channel;
- at least one permeate port fluidically connected to the lower portion of the serpentine channel;
- and
- a first and second reservoir at the first end of the permeate member, wherein the first reservoir is fluidically connected to the at least one retentate port in the retentate member and the second reservoir is fluidically connected to the at least one permeate port in the permeate member;
- a perforated member comprising at least 25,000 perforations disposed under and adjacent to the retentate member;
- a filter disposed under and adjacent to the perforated member and above and adjacent to the permeate member;
- and
- a gasket disposed on top of the reservoir cover of the retentate member, wherein the gasket comprises for each of the first and second reservoir a reservoir access aperture configured to provide fluid access to a reservoir and a pneumatic access aperture configured to provide pneumatic access to a reservoir.
13 more elements
Dependencies are inferred from the claim text.