Polymerase chain reaction container and methods of using the same

• US 5,576,197 A
• Filed: 04/07/1995
• Issued: 11/19/1996
• Est. Priority Date: 04/07/1995
• Status: Expired due to Term

• Current Assignee: Molecular BioProducts Incorporated (Perspecta, Inc.)
• Original Assignee: Molecular BioProducts Incorporated (Perspecta, Inc.)
• Inventors: Arnold, Robert
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US08/418,537
• Time in Patent Office: 592 Days
• Field of Search: 435/6, 435/91.2, 208/20, 208/21
• US Class Current: 435/91.2
• CPC Class Codes: B01L 3/50825 Closing or opening means, corks, bungs closures for containers B65D; means for removing stoppers B67B7/02
• CPC Class Codes: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: C12Q 2547/107 Use of permeable barriers, e.g. waxes
• CPC Class Codes: C12Q 2549/101 Hot start
• CPC Class Codes: G01N 2035/00079 Evaporation covers for slides
• CPC Class Codes: G01N 2035/00287 movable lid/cover for sample or reaction tubes

Abstract

The invention describes a polymerase chain reaction container and methods of using the same that greatly simplify and improve the polymerase chain reaction. Specifically, the invention describes a PCR container comprising a container suitable for PCR with wax attached to the inside surface of the container. The wax is positioned at or above an estimated meniscus position of a PCR mixture. The wax melts at PCR temperatures and covers the surface of the PCR mixture thereby prevents evaporation during thermal cycling. The invention is superior to current methods, in part, because it reduces preparation time, potential sources of contamination and adverse effects on PCR.

Claims

1. A PCR container consisting of the following elements:

a) a container that is suitable for PCR; and

b) has a closed end, an open end distal to the closed end, and a lumen; and

c) a wax attached to the inside surface of the container wherein;

d) the wax is at about the meniscus or above the meniscus position of a PCR mixture added to the container;

e) the wax does not completely occlude the lumen of the container such that the PCR mixture can be added to, the container below the wax without penetrating the wax;

f) the wax is solid at about 40°

C. or below 40°

C. and liquid above 40°

C.;

g) the wax, when liquefied, spontaneously covers the surface of the PCR mixture in the container; and

h) the wax is in an amount that, when liquefied, completely covers the surface of the PCR mixture, with the provision that the inside surface to which the wax is attached is not the bottom of the container and the PCR mixture has not been added to the container.

2. The PCR container of claim 1 wherein the container is a microtube.

3. The PCR container of claim 1 wherein the container is a well in a microtiter plate.

4. The PCR container of claim 1 further comprising the PCR mixture.

5. The PCR container of claim 1 wherein the wax is positioned not less than 0.1 inches from the closed end of the container.

6. The PCR container of claim 5 wherein the container is a conical microtube.

7. The PCR container of claim 5 wherein the amount of wax is about 0.01 to 0.04 g.

8. The PCR container of claim 1 wherein the wax is positioned not more than 0.6 inches from the closed end of the container.

9. A PCR method comprising the following steps:

a) adding a PCR mixture to the PCR container of claim 1;

b) thermally cycling the PCR mixture through PCR temperatures; and

c) completing the PCR method.

10. A hot start PCR method comprising the following steps:

a) adding a PCR mixture lacking an essential reagent to the PCR container of claim 1;

b) heating the PCR container of claim 1 to a PCR temperature;

c) adding the essential reagent lacking in step a to the PCR mixture lacking the essential reagent;

d) thermally cycling the PCR mixture through the PCR temperatures; and

e) completing the hot start PCR method.