Method to evaluate minority variants in a sample

摘要

A method for 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 in step a) to produce fragments; c) attaching an adapter to each end of the fragments created in step b) to produce fragments labeled with adapter d) taking a sample of no more than 10% of the product of step c) and amplifying the fragments labeled with adapter contained in the product sample with the use of one or more primers hybridizing with the adapter 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 readings; f) computationally group the sequence readings for copies of fragments that have the same fragmentation breakpoints and substantially identical sequences to produce groups of readings; g) obtain a consensus sequence for each of the reading groups; h) align the consensus sequences with a reference sequence; and i) identifying a position in the alignment comprising a sequence variation.