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US 11,453,875 B2Granted
Surface-based tagmentation
AssigneeIllumina Cambridge Limited
InventorsNiall Anthony Gormley, Avgousta Ioannou, Rosamond Jackson, Natalie Morrell
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Abstract
Presented herein are methods and compositions surface-based tagmentation. In particular embodiments, methods of preparing an immobilized library of fragmented and tagged DNA molecules on a solid surface are presented. In particular embodiments, the solid surface comprises immobilized transposomes in a dried format, suitable for reconstitution upon contact with liquid, such as a liquid sample.
Claims16
A method of preparing amplified DNA fragments immobilized on a solid support comprising:
- (a) providing a solid support coated with immobilized transposome complexes, wherein the solid support comprises dried tagmentation reagents configured to be reconstituted to form a tagmentation reaction mixture upon contact with a liquid sample, wherein the tagmentation reagents comprise transposome complexes and a tagmentation buffer, wherein the immobilized transposome complexes are prepared by:
- (i) combining purified transposases with transposons to prepare transposome complexes and then immobilizing the transposome complexes to the solid support, or
- (ii) immobilizing transposons to the solid support and then adding purified transposases to prepare immobilized transposome complexes;
- (b) applying a liquid sample comprising target nucleic acid to the solid support under conditions suitable for tagmentation, thereby forming a tagmentation reaction mixture and immobilizing fragments of the target nucleic acid to the solid support;
- (c) washing the solid support to remove any unbound nucleic acids;
- and
- (d) amplifying the immobilized fragments.
4 more elements
Dependencies are inferred from the claim text.