For qualitative detection of calreticulin (CALR) gene mutations by using capillary electrophoresis based on fragment analysis

Kit description

Calreticulin (CALR) is a multifunctional protein that acts as a major Ca(2+)-binding protein in the lumen of the cellular endoplasmic reticulum. It is also found in the nucleus, suggesting to play a role in transcriptional regulation. Calreticulin is encoded by the CALR gene on the chromosome19. Somatic mutations in exon 9 of CALR are the second most prevalent acquired nucleotide changes in Ph-negative myeloproliferative neoplasms (MPNs), except of polycythaemia vera (PV). The two specific mutations are most common, L367fs*46 (type 1 mutation) which represents a 52-bp deletion flanked by 7 base pairs of identical sequence and a K385fs*47(type 2), which results from a 5-bp insertion, and representing an inverse duplication of the five nucleotides preceding the insertion. Overall, these two frameshift mutations types are found in more than 80% of all patients with mutant CALR.

To detect these CALR mutations, one should take the samples of genomic DNA extracted from the bone marrow.

 Optimum range of DNA concentration per reaction is 25-100 ng.

Polymerase chain reaction (PCR)  should be performed by solid-state block PCR thermo cyclers of any firm, but the following PCR conditions are adapted for Bio-Rad thermo cyclers.

Performing of capillary gel electrophoresis is recommended by Life Technologies (3130, 3500, 3500xl) genetic analysers with POP7 polymer.

GeneMapper, Peak Scanner, GeneMarker  programs can serve the  fragment analysis

Ordering information

CALR MutaPrime FA  Kit 4         4 reactions          Cat.No IG-FA2-4         data sheet       instruction         Order free of charge!
CALR MutaPrime FA  Kit 24      24 reactions         Cat.No IG-FA2-24       data sheet       instruction

Related publications:

1.            Nangalia J, Massie CE, Baxter EJ, Nice FL et al. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 2013 Dec 19;369(25):2391-405.
2.            Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H et al. Somatic Mutations of Calreticulin in Myeloproliferative Neoplasms. N Engl J Med. 2013 Dec 19;369(25):2379-90.
3.            Lavi N. Calreticulin mutations in myeloproliferative neoplasms. Rambam Maimonides Med J. 2014 Oct 29;5(4):e0035.