Sample preparation, processing and analysis systems
This disclosure provides an integrated and automated sample-to-answer system that, starting from a sample comprising biological material, generates a genetic profile in less than two hours. In certain embodiments, the biological material is DNA and the genetic profile involves determining alleles at one or a plurality of loci (e.g., genetic loci) of a subject, for example, an STR (short tandem repeat) profile, for example as used in the CODIS system. The system can perform several operations, including (a) extraction and isolation of nucleic acid; (b) amplification of nucleotide sequences at selected loci (e.g., genetic loci); and (c) detection and analysis of amplification product. These operations can be carried out in a system that comprises several integrated modules, including an analyte preparation module; a detection and analysis module and a control module.
1. A microfluidic device comprising:
a) a substrate comprising a microfluidic channel that opens on an inlet;
b) a receptacle comprising:
(i) a compartment comprising a composition comprising one or more reagents;
(ii) a first movable object that seals a top of the compartment; and
(iii) a second movable object that seals an exit port of the compartment and is positioned in a holding chamber in the receptacle;
wherein the receptacle is configured as a vial, and
wherein pressure on the first movable object actuates the second movable object to move out of the holding chamber, thereby providing a fluid flow path around the second movable object, putting the compartment in fluid communication with the inlet through the exit port to allow a fluid in the compartment to flow into the microfluidic channel.
2. The device of claim 1 , wherein the device comprises a plurality of the receptacles each of which comprises a (i) a compartment comprising a composition comprising one or more reagents; (ii) a first movable object that seals a top of the compartment; and (iii) a second movable object that seals an exit port of the compartment and is positioned in a holding chamber in the receptacle.
3. The device of claim 1 , wherein the first movable object is configured as a ball.
4. The device of claim 1 , wherein the second movable object is configured as a ball.
5. The device of claim 1 , wherein the first and second movable objects are configured as balls.
6. The device of claim 1 , wherein the substrate further comprises a reaction chamber that is in fluid communication the one or more microfluidic channel.
7. The device of claim 1 , wherein the composition comprises reagents for performing nucleic acid amplification.
8. The device of claim 2 , wherein the composition comprises reagents for PCR, wherein:
(i) the reagents are optionally stored in a plurality of the receptacles; and/or
(ii) the composition comprises:
one or more pairs of forward and reverse primers for amplifying one or more selected nucleotide sequences, wherein the one or more selected nucleotide sequences optionally comprise one or more selected short tandem repeat (STR) loci;
dye-labeled primers, wherein said dye-labeled primers are optionally labeled with a plurality of dyes;
a polymerase and nucleotide triphosphates, wherein the polymerase optionally comprises a DNA polymerase and the nucleotide triphosphates optionally comprise deoxyribonucleotide triphosphates, wherein the DNA polymerase optionally comprises a Taq polymerase; and/or
a buffering agent and/or a metal salt.
9. The device of claim 1 , wherein:
the composition comprises an STR allelic ladder; and/or
the composition comprises a nucleic acid size standard.
10. The device of claim 1 , wherein the second movable object remains in the receptacle after the compartment is put in fluid communication with the inlet through the exit port.
11. The device of claim 1 , wherein the substrate and the receptacle are formed as a single piece.
12. The device of claim 1 , wherein the receptacle is snapped or press-fitted to the substrate.
13. A microfluidic device comprising:
a) a substrate comprising a microfluidic channel that opens on an inlet;
b) a receptacle comprising:
(i) a compartment comprising a composition comprising one or more reagents;
(ii) a first movable object that seals a top of the compartment; and
(iii) a second movable object that seals an exit port of the compartment and is positioned in a holding chamber in the receptacle, wherein the second movable object is configured as a ball;
wherein pressure on the first movable object actuates the second movable object to move out of the holding chamber, thereby providing a fluid flow path around the second movable object, putting the compartment in fluid communication with the inlet through the exit port to allow a fluid in the compartment to flow into the microfluidic channel.
14. The device of claim 13 , wherein the device comprises a plurality of receptacles each of which comprises (i) a compartment comprising a composition comprising one or more reagents; (ii) a first movable object that seals a top of the compartment; and (iii) a second movable object that seals an exit port of the compartment and is positioned in a holding chamber in the receptacle.
15. The device of claim 13 , wherein the first movable object is configured as a ball.
16. The device of claim 13 , wherein the first and second movable objects are configured as balls.
17. The device of claim 13 , wherein the substrate further comprises a reaction chamber that is in fluid communication the one or more microfluidic channel.
18. The device of claim 13 , wherein the second movable object remains in the receptacle after the compartment is put in fluid communication with the inlet through the exit port.
19. The device of claim 13 , wherein the substrate and the receptacle are formed as a single piece.
20. The device of claim 13 , wherein the receptacle is snapped or press-fitted to the substrate.