Fig. 4.

CALR genotyping and Limit of detection by PCR- ALDA. Dilution series were made by mixing 50% deleted CALR (upper left panel) or inserted CALR (upper right panel) containing DNA samples against wild type DNA. The dilution series were subjected to PCR-ALDA in which only one primer pair is used to unbiasedly amplify wild-type CALR allele and CALR mutant alleles. Subsequently, the reaction mixture was electrophoresed on a 2.5% agarose gel. The 171 bp-CALR-wild-type-allele against the 119 bp (type 1) 52 bp deleted CALR allele or 5 bp TTGTC inserted type − 2 mutation allele or other deleted and inserted alleles. Lower panel- CALR mutations in selected clinical samples: arrows indicate deletions or insertions