Sealing cover and dye compatibility selection

• US 20060029948A1
• Filed: 03/22/2005
• Published: 02/09/2006
• Est. Priority Date: 09/19/2003
• Status: Abandoned Application

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Gunstream, Stephen J., Kinney, Patrick D., Carrillo, Albert L., Lim, Gary, K. Lau, Aldrich N.
• Application Number:
  US11/086,259
• Publication Number:
  US 20060029948A1
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 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 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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 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 9/523 : for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: B01L 9/56 : Means for indicating position of a recipient or sample in an array
• CPC Class Codes: C12Q 1/6816 : characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2531/113 : PCR
• CPC Class Codes: C12Q 2563/173 : staining/intercalating agent, e.g. ethidium bromide
• 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 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/648 : using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates

Abstract

A method of testing compatibility of a detection probe comprising an oligonucleotide and a fluorophore to a composition of a sealing cover, can comprise (a) depositing a quantity of the fluorophore into a plurality of containers; (b) providing a plurality of sealing covers; (c) sealing the plurality of containers with each of the plurality of the sealing covers; (d) exciting the fluorophore; (e) measuring an emission intensity from the fluorophore; and (f) evaluating the emission intensity from the fluorophore to determine the relative compatibility of each of the plurality of sealing covers with the fluorophore.

Claims

1. A method of testing compatibility of a detection probe comprising an oligonucleotide and a fluorophore to a composition of a sealing cover, the method comprising:

depositing a quantity of the fluorophore into a plurality of containers;

providing a plurality of sealing covers wherein each of the plurality of sealing covers comprises a different composition;

sealing the plurality of containers with each of the plurality of the sealing covers;

exciting the fluorophore in each of the plurality of containers;

measuring an emission intensity from the fluorophore in each of the plurality of containers; and

evaluating the emission intensity from the fluorophore in each of the plurality of containers to determine the relative compatibility of each of the plurality of sealing covers with the fluorophore.

2. The method according to claim 1, further comprising controlling a temperature of the plurality of containers during at least one of said exciting the fluorophore in each of the plurality of containers and measuring an emission intensity from the fluorophore in each of the plurality of containers.

3. The method according to claim 2 wherein said controlling a temperature of the plurality of containers during at least one of said exciting the fluorophore in each of the plurality of containers;

and measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises holding the temperature of the plurality of containers constant.

4. The method according to claim 2 wherein said controlling a temperature of the plurality of containers during at least one of said exciting the fluorophore in each of the plurality of containers and measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises cycling the temperature of the plurality of containers.

5. The method according to claim 4 wherein said measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises measuring a plurality of emission intensities from the fluorophore in each of the plurality of containers.

6. The method according to claim 1 wherein said measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises measuring a plurality of emission intensities from the fluorophore in each of the plurality of containers.

7. The method according to claim 6 wherein the cycling the temperature of the plurality of containers further comprises employing a PCR temperature program.

8. A kit for use in polynucleotide amplification, comprising:

a sealing cover;

a pressure sensitive adhesive; and

a detection probe comprising a dye that is compatible with said pressure sensitive adhesive.

9. The kit according to claim 8, further comprising a microplate comprising at least 6,000 wells.

10. The kit according to claim 9 wherein a footprint of the microplate is about 125 cm by about 85 cm.

11. The kit according to claim 9, further comprising the detection probe deposited in at least one of the wells.

12. The kit according to claim 9, further comprising at least one primer deposited in at least one of the wells.

13. The kit according to claim 9, further comprising a non-template control deposited in at least one of the wells.

14. The assembly according to claim 13 wherein the non-template control comprises a fluorescent dye that is compatible with the pressure sensitive adhesive.

15. An apparatus for the analysis of at least one target DNA by PCR, the apparatus comprising:

a microplate comprising at least 400 wells;

a sealing cover comprising a pressure sensitive adhesive and operable to seal a plurality of wells of the microplate;

at least one target DNA in at least one of the 400 wells;

at least one primer set in the at least one of the 400 wells for amplification of the at least one target DNA; and

a detection probe in the at least one of the 400 wells comprising a fluorescent dye that is compatible with the pressure sensitive adhesive.

16. The apparatus according to claim 15 wherein the detection probe further comprises a fluorescent quencher.

17. The apparatus according to claim 16 wherein the fluorescent quencher is compatible with the pressure sensitive adhesive.

18. The apparatus according to claim 15, further comprising a second detection probe in the at least one of the 400 wells comprising a fluorescent dye comprising a second fluorescent dye that is compatible with the pressure sensitive adhesive.

19. The apparatus according to claim 18 wherein the second detection probe is a passive internal reference.

20. The apparatus according to claim 19 wherein the second detection probe further comprises a second fluorescent quencher.

21. The apparatus according to claim 20 wherein the second fluorescent quencher is compatible with the pressure sensitive adhesive.

22. A method of testing compatibility of a detection probe comprising an oligonucleotide and a fluorophore to a composition of a pressure sensitive adhesive, the method comprising:

depositing a quantity of the fluorophore into a plurality of containers;

providing a plurality of sealing covers wherein each of the plurality of sealing covers comprises a different pressure sensitive adhesive;

sealing the plurality of containers with each of the plurality of the sealing covers;

exciting the fluorophore in each of the plurality of containers;

measuring an emission intensity from the fluorophore in each of the plurality of containers; and

evaluating the emission intensity from the fluorophore in each of the plurality of containers to determine the relative compatibility of each of the different pressure sensitive adhesive with the fluorophore.

23. The method according to claim 22, further comprising controlling a temperature of the plurality of containers during at least one of said exciting the fluorophore in each of the plurality of containers and measuring an emission intensity from the fluorophore in each of the plurality of containers.

24. The method according to claim 23 wherein said controlling a temperature of the plurality of containers during at least one of said exciting the fluorophore in each of the plurality of containers and measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises holding the temperature of the plurality of containers constant.

25. The method according to claim 24 wherein said measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises measuring a plurality of emission intensities from the fluorophore in each of the plurality of containers.

26. The method according to claim 23 wherein said controlling a temperature of the plurality of containers during at least one of said exciting the fluorophore in each of the plurality of containers and measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises cycling the temperature of the plurality of containers.

27. The method according to claim 26 wherein said cycling the temperature of the plurality of containers further comprises employing a PCR temperature program.

28. The method according to claim 22 wherein said measuring an emission intensity from the fluorophore in each of the plurality of containers further comprises measuring a plurality of emission intensities from the fluorophore in each of the plurality of containers.