CALR MutaPrime RQ Kit
The test kit is intended for qualitative detection of calreticulin (CALR) gene mutations by using capillary electrophoresis based on fragment analysis
Additional information about the molecular marker
Calreticulin (CALR) is a multifunctional protein that acts as a major Ca(2+)-binding protein in the spaces of 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), but not in 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, thus representing an inverse duplication of the five nucleotides preceding the insert. Overall, these two frameshift mutations are found in more than 80% of all the patients with mutant CALR.
To detect the CALR mutations, the samples of genomic DNA should be extracted from the bone marrow. Optimal range of DNA concentrations per reaction is 25-100 ng.
Polymerase chain reaction (PCR) should be performed by solid-state PCR thermocyclers by any manufacturer, but the following PCR conditions are adapted for Bio-Rad cyclers.
The Life Technologies (3130, 3500, 3500xl) genetic analysers with POP7 polymer are recommended for capillary gel electrophoresis. GeneMapper, Peak Scanner, or GeneMarker software may be used for the fragment analysis
Ordering information:
CALR MutaPrime FA Kit, 24 tests Cat.No FA-2-24
CALR MutaPrime FA Kit, 48 tests Cat.No FA-2-48
Additiional reagents:
Blood Column DNA Kit, 100 extr. Cat.No CDK-100
Price: on request
Reagents are for research use only (RUO)