Use of anticoagulants in the polymerase chain reaction
AssigneeGenedrive Diagnostics Ltd
InventorsShaun Ainsworth, Ben Cobb, Gino Miele
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Abstract
The invention provides a method and kit for preventing inhibition of a thermal cycling reaction by protein coagulation in a sample.
Claims30
A method of performing a thermal cycling reaction comprising:
- a. mixing a sample with i) an anticoagulant and ii) reagents necessary and sufficient for performing a thermal cycling reaction in the sample in the presence of a target nucleic acid, and
- b. incubating the sample under thermal cycling conditions,
- wherein the anticoagulant protects the thermal cycling reaction from inhibition by coagulated proteins in the sample.
The method of any preceding claim, wherein the reagents and anticoagulant are provided in a single solution.
The method of any preceding claim, wherein the anticoagulant comprises MgCl2.
The method of any preceding claim, wherein the anticoagulant comprises Tris, preferably Tris (pH 8.5).
The method of any preceding claim further comprising the step of adding protease to the sample prior to incubating the sample under thermal cycling conditions.
The method of any preceding claim, wherein the sample is subjected to protein denaturing conditions prior to incubating the sample under thermal cycling conditions.
The method of any preceding claim, wherein the thermal cycling reaction is PCR.
The method of any preceding claim, wherein the sample is derived from blood.
The use of arginine as an anticoagulant in a thermal cycling reaction, wherein the arginine prevents coagulation of protein in the sample resulting from heating.
A kit comprising:
- a) a sample tube;
- b) reagents necessary and sufficient to perform a thermal cycling reaction;
- c) arginine.
Dependencies are inferred from the claim text.