Continuous polymerase chain reaction apparatus with multiple temperature stations

• US 6,632,653 B1
• Filed: 12/13/1999
• Issued: 10/14/2003
• Est. Priority Date: 12/08/1997
• Status: Expired due to Term

• Current Assignee: Douglas Scientific LLC
• Original Assignee: Thomas W. Astle
• Inventors: Astle, Thomas W.
• Primary Examiner(s): Whisenant, Ethan
• Assistant Examiner(s): LU, FRANK WEI MIN
• Application Number:
  US09/460,107
• Time in Patent Office: 1,401 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 435/91.5, 435/183, 435/283.1, 435/287.1, 435/287.2, 435/288.4, 436/94, 536/23.1, 536/24.3, 536/24.33, 536/25.3, 422/50, 422/68.1, 422/82.07, 422/82.08
• US Class Current: 435/287.2
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0812 Bands; Tapes
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 3/0265 using valves to interrupt or meter fluid flow, e.g. using solenoids or metering valves
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: B01L 3/527 for a plurality of reagents
• 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: G01N 2035/023 forming cuvettes in situ, e.g. from plastic strip
• CPC Class Codes: G01N 2035/0436 with pre-packaged reagents, i.e. test-packs
• CPC Class Codes: G01N 35/00009 provided with a sample supporting tape, e.g. with absorbent zones

Abstract

In a preferred embodiment, a method of performing a reagent protocol using polymerase chain reaction, including: indexing patterns of reagent wells on a continuous basis through at least one step of reagent addition to the reagent wells; and then indexing the patterns of reagent wells on a continuous basis through a plurality of individual heat transfer stations, whereby at each of the individual heat transfer stations, the patterns of reagent wells are subjected to a unique temperature change to cause one amplification step, with the plurality of individual heat transfer stations providing total amplification required for the protocol.

Claims

1. An apparatus for performing a reagent protocol using polymerase chain reaction, comprising:

(a) means to index patterns of reagent wells on a continuous basis through at least one step of reagent addition to said reagent wells;

(b) means to index said patterns of reagent wells on a continuous basis through a plurality of individual heat transfer stations, whereby at each of said individual heat transfer stations, said patterns of reagent wells are subjected to a unique temperature change to cause one amplification step, with said plurality of individual heat transfer stations providing total amplification required for said protocol; and

(c) means to seal said reagent wells following said at least one step of reagent addition to said wells, wherein said patterns of reagent wells are sealed to provide a liquid tight but peelable seal as provided by pressure sensitive adhesive or heat seal methods.

2. An apparatus for performing a reagent protocol using polymerase chain reaction, comprising:

(a) means to index patterns of reagent wells on a continuous basis through at least one step of reagent addition to said reagent wells;

(b) means to index said patterns of reagent wells on a continuous basis through a plurality of individual heat transfer stations, whereby at each of said individual heat transfer stations, said patterns of reagent wells are subjected to a unique temperature change to cause one amplification step, with said plurality of individual heat transfer stations providing total amplification required for said protocol;

(c) means to seal said reagent wells following said at least one step of reagent addition to said wells; and

(d) separate heat exchanger compartments are clamped to a lower surface of a thermoplastic web containing said patterns of reagent wells to form a liquid tight seal around individual said patterns of reagent wells.

3. An apparatus, as defined in claim 2, further comprising:

means to cause heat exchange fluid to flow through each of said separate heat exchanger compartments for specific time controlled periods.

4. An apparatus for performing a reagent protocol using polymerase chain reaction, comprising:

(a) means to index patterns of reagent wells on a continuous basis through at least one step of reagent addition to said reagent wells;

(b) means to index said patterns of reagent wells on a continuous basis through a plurality of individual heat transfer stations, whereby at each of said individual heat transfer stations, said patterns of reagent wells are subjected to a unique temperature change to cause one amplification step, with said plurality of individual heat transfer stations providing total amplification required for said protocol;

(c) means to seal said reagent wells following said at least one step of reagent addition to said wells; and

(d) means to peel sealing material from a top of said thermoplastic web to provide access to said reagents by a single or multiple well pipettor, said means to peel including a heated pressure roller in contact with said sealing material to apply a line of heat across said thermoplastic web to soften bonding of said sealing material to said thermoplastic web to permit ease of removal by applying tension to said sealing material.