The reciprocal translocation between chromosomes 9 and 22 [t(9;22)(q34;q11), 
Philadelphia chromosome] creates a BCR-ABL1 fusion protein that occurs in 
approximately 95% of cases of chronic myelogenous leukemia (CML), 15% of cases 
of adult acute lymphoblastic leukemia, and 5% of adult cases of acute myeloid 
leukemia. The BCR-ABL1 protein is a constitutively activated tyrosine kinase 
that induces and maintains the neoplastic phenotype in these leukemias. 
PCR-based methods to identify and quantitate the tumor-specific BCR-ABL1 RNA 
have been shown to be an ultrasensitive diagnostic, prognostic, and monitoring 
tool for Philadelphia-positive leukemias. A novel tyrosine kinase inhibitor 
(TKI), imatinib, has been confirmed as an effective targeted treatment in most 
CML patients. However, a significant minority of patients being treated with 
imatinib develop resistance to the drug as evidenced by rising BCR-ABL1 levels. 
The most common mechanism of resistance in these patients is the development of 
mutations in the BCR-ABL1 kinase domain (KD) that abrogate binding of imatinib. 
Although KD mutations are quite heterogeneous, the identification of the exact 
mutation site is clinically important, as some mutations, but not others, can be 
effectively treated with second-generation TKIs. One mutation, T315I, for 
example, renders the leukemia resistant to all first- and second-line TKIs. 
Thus, DNA sequencing of the BCR-ABL1 kinase domain in resistant patients helps 
identify those who may benefit from a change in TKI agents, or those who should 
be considered for other therapeutic measures, such as stem cell transplantation. 
We describe here a method for sequencing the BCR-ABL1 kinase domain in 
peripheral blood or bone marrow of CML patients.