The ipsogen CALR RGQ PCR Kit is a real-time PCR test for the qualitative detection of somatic mutations in the region c.1091_1162 (cDNA annotation) of exon 9 in the CALR gene (GenBank Accession Number CR457070) (3, 4), and also enables the identification of the two major CALR mutations (Type 1 and Type 2). The kit provides reagents to perform seven separate PCR amplification reactions in the same run for the identification of CALR mutations Type 1 and Type 2, and the detection of additional minor variants in genomic DNA (gDNA) extracted from human peripheral whole blood. The turnaround time to execute all tasks, from gDNA extraction (using manual or automated extraction) to data analysis, is less than one working day.
To validate and control the PCR reaction in the presence of human gDNA template, each CALR reaction mix includes primers and probe to amplify an endogenous sequence of the ABL1 housekeeping gene. This control sequence is amplified through a multiplex PCR reaction in all CALR mutant and wild-type DNA. All required components are included in the run to provide accurate and sensitive detection of CALR mutations.
Results analysis and interpretation is automated and simplified with the Rotor-Gene AssayManager v2.1 software.