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US 2015/0087535 A1Published application
Measurement of nucleic acid variants using highly-multiplexed error-suppressed deep sequencing
OwnerAbhijit Ajit Patelinventor
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Abstract
Methods and compositions are disclosed for measuring low-abundance DNA variants from a complex mixture of DNA molecules. Embodiments of the methods allow for extremely sensitive detection and can distinguish true variants from sequencer misreads and PCR misincorporations.
Claims15
A method of measuring nucleic acid variants comprising
- amplifying a low abundance template nucleic acid in a multiplexed polymerase chain reaction (PCR),
- wherein the PCR comprises gene specific primers comprising a) a barcode;
- and b) a molecular lineage tag (MLT),
- wherein the gene specific primers are specific for a mutation prone region.
1 more element
A method of primer extension comprising targeted early barcoding.
A method of distinguishing true nucleic acid variants from nucleotide misincorporations during PCR comprising
- a) amplifying template nucleic acids with primers comprising MLTs, wherein there is a diversity of MLT sequences and a MLT sequence can be assigned to more than one template molecule;
- and
- b) identifying misincorporation errors wherein sampled clonal sequences are several-fold greater than tagged template copies.
Dependencies are inferred from the claim text.