Method for isolating nucleic acid mutations
AssigneeBoreal Genomics, Inc.
InventorsAndrea Marziali, Joel Pel
Life of this patent
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Abstract
The invention includes methods and apparatus for isolating and recovering mutations, especially rare and unknown mutations, without amplifying the sample. In particular, using the disclosed methods, it is possible to separate heteroduplexed nucleic acid strand pair from homoduplexed nucleic acid strand pairs having similar sequences and being at a much higher concentration. Once the heteroduplexed nucleic acids are isolated and recovered, it is straightforward to analyze the sequences of the heteroduplexed nucleic acids, e.g., using sequencing or hybrid assays.
Claims21
A method for separating a mixture of homoduplexed and heteroduplexed nucleic acids, comprising;
- loading a sample comprising a mixture of homoduplexed and heteroduplexed nucleic acids on a separation medium;
- applying a time-varying driving field to the separation medium;
- and
- applying a time-varying mobility varying field to the separation medium,
- thereby causing the homoduplexed nucleic acids to be separated from the heteroduplexed nucleic acids.
A method for determining a mutation in a nucleic acid sample, comprising:
- amplifying a plurality of non-identical nucleic acids in a sample to create a plurality of non-identical amplicons;
- denaturing and reannealing the non-identical amplicons to produce a sample comprising a mixture of homoduplexed and heteroduplexed nucleic acids;
- loading the sample comprising a mixture of homoduplexed and heteroduplexed nucleic acids on a separation medium;
- applying both a time-varying driving field and a time-varying mobility varying field to the separation medium, thereby causing the homoduplexed nucleic acids to be separated from the heteroduplexed nucleic acids;
- and
- recovering the heteroduplexed nucleic acids.
Dependencies are inferred from the claim text.