Method of nucleic acid amplification
AssigneeIllumina, Inc.
InventorsEric H. Kawashima, Laurent Farinelli, Pascal Mayer
Life of this patent
Estimating the term…
Abstract
A nucleic acid molecule can be annealed to an appropriate immobilized primer. The primer can then be extended and the molecule and the primer can be separated from one another. The extended primer can then be annealed to another immobilized primer and the other primer can be extended. Both extended primers can then be separated from one another and can be used to provide further extended primers. The process can be repeated to provide amplified, immobilized nucleic acid molecules. These can be used for many different purposes, including sequencing, screening, diagnosis, in situ nucleic acid synthesis, monitoring gene expression, nucleic acid fingerprinting, etc.
Claims31
(canceled)
A process that can be used to identify in a nucleic acid sample the presence or absence of nucleic acid sequence differences, wherein each said difference is with respect to a reference sequence, the process comprising:
- a. fragmenting nucleic acids in said sample;
- b. ligating adapter sequences to the nucleic acid fragments generated in step a;
- c. replicating said adapter linked nucleic acid fragments of step b;
- d. binding said replicated and adapter ligated nucleic acid fragments to a solid support by hybridizing oligonucleotides attached to the solid support to the adapter sequences;
- and
- e. identifying nucleic acid sequences within said replicated and solid support bound nucleic acid fragments by hybridizing primers to the bound and replicated fragments and extending the primers while the primers are hybridized to the replicated and solid support bound fragments.
A process to identify in a nucleic acid sample the presence or absence of nucleic acid sequence differences, wherein each said difference is with respect to one or more reference sequences, the process comprising:
- a. contacting a nucleic acid sample, or molecules replicated from said nucleic acid sample, with nucleic acid molecules comprising sequence-specific binding activity under conditions which permit binding;
- b. linking said molecules comprising sequence-specific binding activity to one another when said molecules bind to the nucleic acid sample, or a replica thereof, under conditions which permit said molecules to be linked;
- c. replicating the linked molecules of step b;
- d. binding the replicated molecules of step c to a solid support to form bound replicated molecules;
- e. identifying nucleic acid sequences within said replicated molecules.
A process to identify in a nucleic acid sample the presence or absence of nucleic acid sequence differences, wherein each said difference is with respect to one or more reference sequences, the process comprising:
- a. contacting a nucleic acid sample, or molecules replicated from said nucleic acid sample, with nucleic acid molecules comprising sequence-specific binding activity under conditions which permit binding;
- b. linking said molecules comprising sequence-specific binding activity to one another when said molecules bind to the nucleic acid sample, or a replica thereof, under conditions which permit said molecules to be linked;
- c. binding the linked molecules to a solid support and replicating said bound molecules;
- d. identifying nucleic acid sequences within said replicated molecules.
Dependencies are inferred from the claim text.