Method for Evaluating Minority Variants in a Sample

摘要

A method of evaluating a sequence variation in a sample is provided. In some embodiments, the method may involve: amplifying a nucleic acid product from an initial sample; fragmenting an amount of the nucleic acid product to produce fragments; attaching an adaptor to each end of the fragments to produce adaptor-tagged fragments; sampling no more than 10% of the tagged fragments and amplifying them; sequencing at least some of the copies of the fragments to produce a plurality of sequence reads; grouping sequence reads for copies of fragments that have the same fragmentation breakpoints; deriving a consensus sequence for each of the read groups; and aligning the consensus sequences with a reference sequence.

主权项

1. A method of evaluating a sequence variation in a sample, comprising:
a) amplifying a nucleic acid product from an initial sample; b) fragmenting an amount of the nucleic acid product produced by step a) to produce fragments; c) attaching an adaptor to each end of the fragments created in step b) to produce adaptor-tagged fragments d) sampling no more than 10% of the product of step c) and amplifying the adaptor-tagged fragments contained in the sampled product using one or more primers that hybridize to the adaptor to produce copies of the fragments e) sequencing at least some of the copies of the fragments produced in d) to produce a plurality of sequence reads; f) computationally grouping sequence reads for copies of fragments that have the same fragmentation breakpoints and substantially identical sequences to produce read groups; g) deriving a consensus sequence for each of the read groups; h) aligning the consensus sequences with a reference sequence; and i) identifying a position in the alignment that comprises a sequence variation.