Automated process for preparing and amplifying a target nucleic acid sequence
View Patent ↗An automated process for preparing and amplifying a target nucleic acid sequence within a stand-alone unit.
1. A process for preparing and amplifying a target sequence contained within a target nucleic acid present in a fluid sample, the process comprising performing in a stand-alone unit the ordered and automated steps of:
a) immobilizing the target nucleic acid on a solid support;
b) separating other material present in the fluid sample from the target nucleic acid immobilized in step a);
c) washing the solid support one or more times with a wash solution; and
d) amplifying the target sequence in a receptacle containing the target nucleic acid and amplification reagents provided thereto, the receptacle being formed to have an open top end and a closed bottom end wherein the stand-alone unit comprises first and second stations, and wherein steps a)–c) are performed at the first station and step d) is performed at the second station.
2. The process of claim 1 wherein the first and second stations are contained within a housing.
3. The process of claim 2 , wherein the first and second stations are located on a processing deck.
4. The process of claim 2 further comprising performing in the stand-alone unit the automated step of transferring the target nucleic acid from the first station to the second station.
5. The process of claim 4 , wherein the transferring step is performed using a transport mechanism rotatable about an axis of rotation.
6. The process of clam 1 , wherein the solid support comprises a magnetically responsive particle.
7. The process of claim 6 , wherein the separating step comprises subjecting the solid support to a magnetic field and aspirating the other material present in the fluid sample.
8. The process of claim 7 , wherein the other material present in the fluid sample includes non-target nucleic acid.
9. The process of claim 1 , wherein a robotic pipettor is used to combine the target nucleic acid and the reagents in step d).
10. The process of claim 1 further comprising performing in the stand-alone unit the automated step of exposing the target sequence or its complement to a probe under conditions sufficient to permit the probe to hybridize to the target sequence or its complement.
11. The process of claim 10 further comprising performing in the stand-alone unit the automated step of detecting the presence of the probe hybridized to the target sequence or its complement.
12. The process of claim 1 , wherein the receptacle is a tube.
13. The process of claim 1 , wherein the receptacle is a molded plastic.
14. The process of claim 1 , wherein the receptacle is one of an integrally formed plurality of receptacles.
15. A process for preparing and amplifying a target sequence contained within a target nucleic acid present in a fluid sample, the process comprising performing in a stand-alone unit the ordered and automated steps of:
a) immobilizing the target nucleic acid on a solid support;
b) separating other material present in the fluid sample from the target nucleic acid immobilized in step a);
c) washing the solid support one or more times with a wash solution; and
d) amplifying the target sequence in a tubular receptacle containing the target nucleic acid and amplification reagents provided thereto, wherein steps a)–d) are performed in the stand-alone unit.
16. The process of clam 15 , wherein the solid support comprises a magnetically responsive particle.
17. The process of claim 16 , wherein the separating step comprises subjecting the solid support to a magnetic field and aspirating the other material present in the fluid sample.
18. The process of claim 15 , wherein the other material present in the fluid sample includes non-target nucleic acid.
19. The process of claim 15 , wherein a robotic pipettor is used to combine the target nucleic acid and the reagents in step d).
20. The process of claim 15 further comprising performing in the stand-alone unit the automated step of exposing the target sequence or its complement to a probe under conditions sufficient to permit the probe to hybridize to the target sequence or its complement.
21. The process of claim 20 further comprising performing in the stand-alone unit the automated step of detecting the presence of the probe hybridized to the target sequence or its complement.
22. The process of claim 15 , wherein the receptacle is a molded plastic.
23. The process of claim 15 , wherein the receptacle is one of an integrally formed plurality of receptacles.
24. A process for preparing and amplifying a target sequence contained within a target nucleic acid present in a fluid sample, the process comprising performing within a housing of a stand-alone unit the ordered and automated steps of:
a) at a first station contained within the housing, immobilizing the target nucleic acid on a magnetically responsive particle; subjecting the magnetically responsive particle to a magnetic field and aspirating other material present in the fluid sample from the immobilized target nucleic acid; and washing the magnetically responsive particle one or more times with a wash solution;
b) transferring the target nucleic acid to a second station contained within the housing; and
c) at the second station, amplifying the target sequence in a receptacle containing the target nucleic acid and amplification reagents provided thereto, wherein the receptacle is one of an integrally formed plurality of receptacles.
25. The process of claim 24 , wherein the first and second stations are located on a processing deck.
26. The process of claim 24 , wherein the transferring step is performed using a transport mechanism rotatable about an axis of rotation.
27. The process of claim 24 , wherein the other material present in the fluid sample includes non-target nucleic acid.
28. The process of claim 24 , wherein a robotic pipettor is used to combine the target nucleic acid and the reagents in step c).
29. The process of claim 24 further comprising performing within the housing the automated step of exposing the target sequence or its complement to a probe under conditions sufficient to permit the probe to hybridize to the target sequence or its complement.
30. The process of claim 29 further comprising performing within the housing the automated step of detecting the presence of the probe hybridized to the target sequence or its complement.
31. The process of claim 24 , wherein each of the receptacles is a tube.
32. The process of claim 24 , wherein the integrally formed plurality of receptacles is a molded plastic.