Integrated sample preparation systems and stabilized enzyme mixtures
AssigneeIbis Biosciences, Inc.
InventorsMark Eshoo, John Picuri
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Abstract
The present invention provides integrated sample preparation systems and stabilized enzyme mixtures. In particular, the present invention provides microfluidic cards configured for processing a sample and generating DNA libraries that are suitable for use in sequencing methods (e.g., next generation sequencing methods) or other suitable nucleic acid analysis methods. The present invention also provides stabilized enzyme mixtures containing an enzyme (e.g., an enzyme used in whole genome amplification), BSA, and a sugar. Such enzyme mixtures may be lyophilized and stored at room temperature without significant loss of enzyme activity for months.
Claims14
A microfluidic card comprising:
- a) a loading port configured for introduction of a biological sample;
- b) a nucleic acid extraction sub-circuit wherein said nucleic acid extraction sub-circuit comprises:
- i) a nucleic acid extraction component configured to bind nucleic acids present in said biological sample;
- ii) a wash buffer;
- iii) an elution buffer;
- and
- iv) a component configured for moving said elution buffer over said nucleic acid extraction component to generate a mixture of extracted nucleic acids;
- and
- c) an amplification sub-circuit, wherein said amplification sub-circuit comprises an enzyme mixture, wherein said enzyme mixture comprises at least one amplification-related enzyme useful for performing amplification on said extracted nucleic acid to generate amplified nucleic acid,
- d) a fragmentation sub-circuit, wherein said fragmentation sub-circuit comprises:
- i) a reagent mixture configured for digesting nucleic acid to generate fragmented nucleic acid;
- and/or
- ii) a fragmentation component configured for mechanically fragmenting nucleic acid to generate fragmented nucleic acid;
- and
- iii) at least one type of enzyme configured for polishing the ends of said fragmented nucleic acid;
- and
- e) a linker sub-circuit, wherein said linker ligation sub-circuit comprises:
- i) nucleic acid linkers configured for use in sequencing methods;
- ii) a ligation enzyme mixture configured for ligating said nucleic acid linkers to nucleic acid to generate a nucleic acid sequencing library;
- and
- iii) at least one type of enzyme configured for polishing the ends of said nucleic acid sequencing library;
- and
- f) a purification sub-circuit, wherein said purification sub-circuit comprises a nucleic acid purification component.
Dependencies are inferred from the claim text.