Methods and kits for detecting gene mutations
AssigneeShuwen Biotech Co. Ltd.
InventorsXingmin Li, Huanhuan Yang
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Abstract
The invention relates to methods, kits and uses thereof for detecting gene mutations. The method comprises the step of: providing a mixture of nucleic acid molecules comprising both a genomic DNA and a RNA molecule derived from said genomic DNA, performing a reverse transcription reaction with a reverse transcriptase to provide cDNA molecules from said RNA molecule in said mixture; and performing PCR using both said genomic DNA and said cDNA molecules as templates. This method can simultaneously detect a trace amount of a mutation in both DNA and RNA from a whole blood sample using a small amount of sample. At the same time, by adjusting the reaction system, the detection limit of a mutant in a background of wild-type molecules can reach 0.01% or lower.
Claims6
A method for detecting a mutation in a biological sample, comprising:
- providing a mixture of nucleic acid molecules comprising both a genomic DNA and a RNA molecule derived from said genomic DNA, both said genomic DNA and said RNA molecule being extracted from a human whole blood sample from a cancer patient;
- performing a reverse transcription reaction with a reverse transcriptase to provide cDNA molecules from said RNA molecule in said mixture;
- and
- performing PCR using both said genomic DNA and said cDNA molecules as templates,
- determining whether a mutation is present based on the result of the PCR.
A PCR reaction mixture, comprising:
- a mixture of nucleic acid molecules comprising both a genomic DNA of a human gene and a RNA molecule derived from said genomic DNA, both extracted from a human whole blood sample from a cancer patient, in admixture with;
- a reverse transcriptase,
- a Taq DNA polymerase, and
- a pair of PCR primers,
- wherein one primer of said pair of PCR primers is an ARMS primer.
A kit for detecting a gene mutation in a biological sample, comprising:
- a first primer pair capable of amplifying by PCR a DNA sequence of a gene;
- a RT primer for priming reverse transcription from a RNA transcript of said DNA sequence to provide a cDNA;
- a second primer pair capable of amplifying by PCR all or part of said cDNA sequence;
- and
- an enzyme mixture having both a reverse transcriptase and a Taq polymerase,
- wherein one primer of said second primer pair is an ARMS primer.
Dependencies are inferred from the claim text.