Method of primer-extension preamplification PCR
AssigneeRoche Molecular Systems, Inc.
InventorsWolfgang Dietmaier, Josef Ruschoff
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Abstract
The invention provides a method for the amplification of nucleic acid fragments from a sample that comprises two or three thermocyclic amplification reactions, whereby completely randomized primers are used in the first amplification reaction and specific primers are used in the second amplification reaction, characterized in that, to amplify the DNA, a mixture of at least two DNA polymerases is used, at least one of which possesses proofreading activity. Using this method, DNA from single cells or cell clones with a low cell count can be used in mutation analysis.
Claims4
A method for the amplification of nucleic acid fragments from a sample, said method comprising first and second thermocyclic amplification reactions, wherein said first amplification reaction is carried out using completely randomized primers, said second amplification reaction is carried out using specific primers, and said first and second amplification reactions are carried out using a mixture of at least two DNA polymerases, at least one of which possesses 3′-5′ exonuclease activity.
Dependencies are inferred from the claim text.