Normalization of data using controls

• US 7,417,726 B2
• Filed: 03/22/2005
• Issued: 08/26/2008
• Est. Priority Date: 09/19/2003
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Oldham, Mark F., Harding, Ian A., Karger, Achim, Kao, H. Pin, Ostadan, Omead, Young, Greg
• Primary Examiner(s): EVANS, FANNIE L
• Application Number:
  US11/087,103
• Publication Number:
  US 20060024831A1
• Time in Patent Office: 1,253 Days
• Field of Search: 436/8, 356/243.1, 356/243.5
• US Class Current: 356/243.1
• CPC Class Codes: B01L 2200/021 Adjust spacings in an array of wells, pipettes or holders, format transfer between arrays of different size or geometry
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/022 Transponder chips
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0851 Bottom walls
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: B01L 3/563 Joints or fittings in general F16L; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 9/50 : Clamping means, tongs in general F16B2/06
• CPC Class Codes: B01L 9/523 : for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: G01N 2035/00366 : Several different temperatures used
• CPC Class Codes: G01N 21/274 : Calibration, base line adjustment, drift correction
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates
• CPC Class Codes: H04N 25/63 : applied to dark current
• CPC Class Codes: H04N 25/633 : by using optical black pixels
• CPC Class Codes: H04N 25/68 : applied to defects
• CPC Class Codes: Y10T 436/10 : Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing

Abstract

An apparatus for normalizing a PCR instrument, can comprise a microplate comprising at least 6,000 wells and a system of dyes at known concentrations in a plurality of the at least 6,000 wells. A method for normalizing a system can comprise (a) providing a microplate comprising at least 6,000 wells and a system of dyes; (b) exciting at least one dye; (c) detecting an emission output for the at least one dye; (d) determining if the emission output is in an acceptable range; and (e) adjusting the system so that the emission output is in the acceptable range.

Claims

1. A calibration apparatus for calibrating an instrument, the instrument capable of polymerase chain reaction, the apparatus comprising:

a microplate comprising at least 400 wells; and

a plurality of the wells each having a mixture of dyes at known concentrations spatially distributed throughout the microplate.

2. The apparatus according to claim 1, further comprising a sealing cover coupled to the microplate.

3. The apparatus according to claim 1 wherein the microplate has dimensions of about 127 cm by about 85 cm.

4. The apparatus according to claim 1 wherein the at least 400 wells is 6,144 wells.

5. The apparatus according to claim 1 wherein the plurality of the wells each having a mixture of dyes comprises a series dilution.

6. A method for calibrating an instrument, the method comprising:

providing a first instrument;

placing a microplate comprising at least 6,000 wells and a plurality of dyes in the first instrument;

exciting at least one dye of the plurality of dyes;

detecting an emission output from the at least one dye;

determining if the emission output from the at least one dye is within a predetermined range;

calibrating the first instrument so that the emission output from the at least one dye is within the predetermined range;

providing a second instrument;

placing the microplate comprising at least 6,000 wells and a plurality of dyes in the second instrument;

exciting at least one dye of the plurality of dyes;

detecting an emission output from the at least one dye;

determining if the emission output from the at least one dye is within a predetermined range; and

calibrating the second instrument so that the emission output from the at least one dye is within the predetermined range.

7. The method according to claim 6, further comprising cycling a temperature of the microplate.

8. A method according to claim 6, further comprising:

exciting a second dye of the plurality of dyes;

detecting an emission output from the second dye;

determining if the emission output from the second dye is within a predetermined range; and

calibrating the first instrument so that the emission output from the second dye is within the predetermined range.

9. The method according to claim 6 wherein the plurality of dyes comprises a series dilution.

10. The method according to claim 6 wherein the calibrating the first instrument so that the emission output from the at least one dye is within the predetermined range further comprises adjusting an excitation source of the system.

11. The method according to claim 6 wherein the calibrating the first instrument so that the emission output from the at least one dye is within the predetermined range further comprises adjusting a detector.

12. The method according to claim 6 wherein the calibrating the first instrument so that the emission output from the at least one dye is within the predetermined range further comprises adjusting an optical component.

13. The method according to claim 6 wherein the calibrating the first instrument so that the emission output from the at least one dye is within the predetermined range further comprises adjusting an electronic component.

14. A method for normalizing a first PCR system as compared to a second PCR system, the method comprising:

providing a first PCR system;

placing a microplate comprising at least 400 wells and a plurality of dyes in at least one of the at least 400 wells into the first PCR system;

exciting at least one dye of the plurality of dyes;

detecting an emission output from the at least one dye;

evaluating an adjustment to the first PCR system based on the emission output for the at least one dye so that the first PCR system is normalized to the second PCR system; and

adjusting the first PCR system operably normalizing it to the second PCR system.

15. The method according to claim 14, further comprising:

exciting a second dye;

detecting an emission output of the second dye;

evaluating an adjustment to the first PCR system based on the emission output for the second dye so that the first PCR system is normalized to the second PCR system; and

adjusting the first PCR system operably normalizing it to the second PCR system.

16. The method according to claim 14 wherein the adjusting the first PCR system operably normalizing it to the second PCR system, further comprising adjusting an optical component of the first PCR system.

17. The method according to claim 14 wherein the adjusting the first PCR system operably normalizing it to the second PCR system, further comprising adjusting an electronic component of the PCR system.

18. The method according to claim 14, further comprising cycling a temperature of the microplate.

19. The method according to claim 14, further comprising performing a PCR program.

20. The method according to claim 14, further comprising detecting an emission output of the at least one dye at each step of a PCR program.

21. The method according to claim 14 wherein the plurality of dyes comprise a series dilution.