Methods for amplifying a nucleic acid

Abstract

Provided is a method for amplifying a nucleic acid which is suspected to be present in a sample in a low copy number comprises the following steps: i. providing a first reaction mixture in a reaction chamber wherein the first reaction mixture comprises a first portion of the sample; ii. subjecting the first reaction mixture in the reaction chamber to conditions allowing a pre-amplification of the nucleic acid in order to obtain a first pre-amplification mixture; iii. providing a second reaction mixture in the reaction chamber wherein the second reaction mixture comprises a subset of the first pre-amplification mixture and a second portion of the sample; iv. subjecting the second reaction mixture to conditions allowing a pre-amplification of the nucleic acid in order to obtain a second pre-amplification mixture; v. optionally, repeating steps iii) and iv) a number N additional times in order to obtain an (N+2) pre-amplification mixture; vi. subjecting the second pre-amplification mixture obtained in step (iv) or the (N+2) pre-amplification mixture obtained in step (v) to conditions allowing an amplification of the nucleic acid in order to obtain an amplification mixture, wherein the sample has a total sample volume which exceeds a volume capacity of the reaction chamber.

Dependencies are inferred from the claim text.