Polymerase chain reaction (PCR) amplification for the detection of the FLT3 ITD and FLT3 TKD mutations was performed on DNA isolated from the patient sample. Fluorescently labeled primers amplified the sequences of interest. The TKD PCR product was cut with the EcoRV restriction enzyme. The ITD and the digested TKD products were run on the ABI 3500x1 genetic analyzer and their sizes were determined. The ITD wild type produces a fragment that is approximately 327 +/-1 bp, while the presence of an insertion produces a fragment that is approximately >=330 bp. This assay has an analytical sensitivity of 5 mutation containing cells in 100 total cells. This assay does not detect ITD mutations greater than approximately 420 bp in size.
3 - 7 days
5 - 9 days
Laboratory for Personalized Molecular Medicine