METHOD TO INCREASE SENSITIVITY OF NEXT GENERATION SEQUENCING

摘要

A method for detecting a low-occurrence mutation in isolated DNA adds a blocking probe to reagents during amplification of the isolated DNA. The blocking probe is an oligonucleotide complementary to wild-type DNA corresponding to the sample. The blocking probe spans a site of a suspected mutation within a region of interest in the isolated DNA. After amplification, fragments of the amplified DNA is sequenced using next generating sequencing and an output is generated to display the sequenced fragments. In some embodiments, the blocking probe is locked nucleic acid (LNA).

主权项

1. A method of detecting a low-occurrence mutation in a sample from a patient, the method comprising:
isolating DNA from the sample; adding a blocking probe to the isolated DNA, the blocking probe comprising an oligonucleotide complementary to wild-type DNA corresponding to the sample, the blocking probe adapted to span a site of a suspected mutation within a region of interest in the isolated DNA; amplifying the isolated DNA; sequencing fragments of the amplified DNA using next generating sequencing; and generating an output corresponding to the sequenced fragments.