Mutation identification method

A method for identifying the position of a genetic mutation, comprising:

• a) determining an ascertained nucleic acid (ANA) region comprising the genetic mutation;
• b) performing polymerase chain reaction (PCR) with primers to generate a PCR product, wherein the PCR template is DNA extracted from a biological sample obtained from a subject, and the primers are:
• b1) a degenerate primer, and
• b2) a specific primer:
• hybridizing specifically within the ANA region, at a distance of 0.2-100 Kb from any end of the ANA region, if the genetic mutation comprises a gain of genetic material, a translocation or a gene fusion, and extending by PCR out of the ANA region, or
• hybridizing specifically outside the ANA region, at a distance of 0.2-100 Kb from any end of the ANA region, if the mutation comprises a loss of genetic material, and extending by PCR into the ANA region,
• and wherein in the PCR, the hybridizing comprises two consecutive hybridizations, a first hybridization at a hybridization temperature equal to the Tm of the specific primer ±10° C., and a second hybridization at a hybridization temperature equal to the Tm of the degenerate primer ±5° C.;
• c) sequencing the PCR product obtained in step b) to obtain a sequence;
• and
• d) aligning the sequence obtained in step c) with a reference genome for identifying the position of the genetic mutation.