Patent ID: 8470261

Claim:
A microfluidic card comprising: a) a loading port configured for introduction of a biological sample; and b) a lysis sub-circuit operably linked to said loading port, wherein said lysis sub-circuit comprises: i) a lysis chamber; and ii) a lysis buffer wherein said lysis sub-circuit is configured to lyse said biological sample with said lysis buffer to generate a lysed sample; c) a nucleic acid extraction sub-circuit operably linked to said lysis sub-circuit wherein said nucleic acid extraction sub-circuit comprises: i) a nucleic acid extraction component configured to bind nucleic acids present in said lysed sample; ii) a wash buffer; iii) an elution buffer; and iv) a pump component configured for pumping said elution buffer over said nucleic acid extraction component to generate a mixture of extracted nucleic acids; and d) an amplification sub-circuit operably linked to said nucleic acid extraction sub-circuit, wherein said amplification sub-circuit comprises a stabilized enzyme mixture, wherein said stabilized enzyme mixture comprises at least one amplification-related enzyme useful for performing amplification on said extracted nucleic acid to generate amplified nucleic acid, e) a fragmentation sub-circuit operably linked to said amplification sub-circuit, wherein said fragmentation sub-circuit comprises: i) a reagent mixture configured for digesting said amplified nucleic acid to generate fragmented nucleic acid; and/or ii) a fragmentation component configured for mechanically fragmenting said amplified 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 f) a linker ligation sub-circuit operably linked to said fragmentation 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 said fragmented 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 g) a purification sub-circuit operably linked to said linker ligation sub-circuit wherein said purification sub-circuit comprises a nucleic acid purification component.