Chemistry: molecular biology and microbiology – Apparatus – Bioreactor
Reexamination Certificate
2002-01-04
2003-10-14
Redding, David A. (Department: 1744)
Chemistry: molecular biology and microbiology
Apparatus
Bioreactor
C435S288400, C435S309100, C435S395000, C435S287300
Reexamination Certificate
active
06632660
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to devices and methods for use in biochemical procedures, and to solid supports upon which one or more species can be immobilized.
BACKGROUND OF THE INVENTION
There has been a desire in recent years to develop methods for reversibly attaching biochemicals such as terminators or primers to a solid support. Various means have been considered for capturing and releasing nucleotides from a support, and ultimately delivering them to a specific site. Applications include hybridizing individual biological samples to mixed terminators that have been placed in the wells of microtiter plates, and then eluting the labeled nucleotide chains from their support for use in multiple sequencing reactions. Multiplex sequencing has the potential to significantly decrease sample handling and associated consumable costs for sample preparation for large-scale sequencing and genetic analysis projects. In order to selectively retrieve or “fish out” separate samples, the use of modified solid supports has been investigated.
SUMMARY OF THE INVENTION
Aspects of the present invention relate to, among other things, apparatus and methods providing an array of binding sites within the wells of a microplate (e.g., a standard-format, 96- 384-well plate). According to various embodiments, such binding sites comprise an array of petal-like members adapted for insertion into a corresponding array of reaction wells. Further aspects of the invention relate to, among other things, apparatus and methods facilitating the release of labeled monomers from a support, also within the microplate format.
Various embodiments provide an analyte-manipulation apparatus. The apparatus can include, for example, a plurality of wells defining an array, wherein each of the wells includes a rim defining an opening at an upper end thereof, with the openings being disposed within a first plane. The apparatus can further include a sheet-like support including a plurality of petal-like members integrally formed therein at positions corresponding to the wells of the array, with the support being disposed along a second plane above and substantially parallel to the first plane, and with at least one of the petal-like members being positioned near each one of the openings. In various embodiments, each of the petal-like members is movable between (i) a first position, substantially within the second plane, and (ii) a second position, at least partially disposed outside of the second plane and extending at least partially into a nearby well via a respective opening. The apparatus can further include a platen including a major surface facing the support, and a plurality of ring-like projections extending outwardly from the major surface, with the platen being adapted for movement toward and away from the support, whereby upon moving the platen toward the support, the projections can pressingly engage the petal-like members, thereby deflecting the petal-like members from the first position to the second position.
In various embodiments, the apparatus further includes a die plate disposed between the support and the rims, with the die plate including an array of apertures extending therethrough, and with each of the apertures being disposed at a position corresponding to a respective one of the wells of the array.
According to various embodiments, the platen and each of the ring-like projections defines a passage extending longitudinally through each ring-like projection and through the platen. Such passage can be quite useful. For example, with the petal-like members being deflected into their respective wells, an instrument such as a pipette or the like can be inserted through the passage to access the interior region of any one or more of the wells. For example, sample and/or reagent can be deposited into one or more selected wells, e.g., using such an instrument, via such passage. In addition, or in the alternative, sample and/or reagent can be withdrawn from one or more selected wells, e.g., using such an instrument, via such passage.
In various embodiments, at least a portion of the petal-like members can be chemically treated.
According to various embodiments, one or more of the petal-like members can include one or more biochemicals immobilized thereon. Such biochemicals can include, for example, one or more nucleic acids. In various embodiments, such biochemicals comprise one or more DNA-sequencing reagents, such as terminators, primers, or a combination thereof.
According to various embodiments, each support is a single-layer film or membrane material.
In various embodiments, the petal-like members are resiliently deformable, tending to return to the first position after having been deflected therefrom.
According to various embodiments, each support includes one or more registration features (e.g., one or more slots formed in each of the sheets) facilitating alignment of the supports with respect to the microplate.
In various embodiments, each of the ring-like projection tapers in a direction away from the major surface.
Various embodiments provide an analyte-manipulation apparatus including a plurality of wells defining an array, with each of the wells including an opening at an upper end thereof. The apparatus can further include a stack of sheet-like supports disposed above the openings, with each support of the stack including a plurality of petal-like members integrally formed therein, and with each petal-like member of each support being disposed at a position corresponding to a respective one of the wells of the array. In various embodiments, each of the petal-like members is movable between (i) a first position, outside of a corresponding respective well, and (ii) a second position, extending at least partially into such corresponding respective well. The apparatus can further include a platen including a major surface facing the support, and a plurality of ring-like projections extending outwardly from the major surface, with the platen being adapted for movement toward and away from the support, whereby upon moving the platen toward the support, the projections can pressingly engage the petal-like members, thereby deflecting the petal-like members from the first position to the second position.
According to various embodiments, the stack includes at least three (e.g., 3, 4, 5, 6, 7, 8, 9, 10, or more) of the supports.
In various embodiments, one or more of the petal-like members includes one or more biochemicals (e.g., nucleic acids) immobilized thereon.
According to various embodiments, each support is a single-layer film or membrane material.
According to various embodiments, the platen and each of the ring-like projections defines a passage extending longitudinally through each ring-like projection and through the platen. Such passage can be quite useful. For example, with the petal-like members being deflected to the second position, an instrument such as a pipette or the like can be inserted through the passage to access the interior region of any one or more of the wells. For example, sample and/or reagent can be deposited into one or more selected wells, e.g., using such an instrument, via such passage. In addition, or in the alternative, sample and/or reagent can be withdrawn from one or more selected wells, e.g., using such an instrument, via such passage.
Various embodiments provide a method for biochemical interactions.
In certain embodiments, such a method includes:
immobilizing one or more selected biochemicals on a plurality of petal-like members, wherein the petal-like members are disposed in an array on a support;
introducing one or more reagents into a plurality of wells, wherein the wells are disposed in an array corresponding to the array of petal-like members;
positioning the petal-like members above the plurality of wells, so that each petal-like member is situated above a corresponding one of the plurality of wells;
pressingly engaging the petal-like members from a side opposite that of the wells, so that the petal-like members are simultaneously mov
Applera Corporation
Frazier Jeffery D.
Redding David A.
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