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CAT #: 21010181

Hypermutation Mix 2 v2.0 - 6FAM for ABI Fluorescence Detection

Product Use

Hypermutation Mix 2 v2.0 – 6FAM for ABI Fluorescence Detection is a Research Use Only master mix that can be used to identify clonal rearrangements of the immunoglobulin heavy (IGH) chain gene and determine the somatic mutation status of the variable (V) gene.

Product Details

  • Description

    Rearrangements of the antigen receptor genes occur during ontogeny in B and T lymphocytes.  These gene rearrangements are unique in length and sequence for each cell.  Therefore, polymerase chain reaction (PCR) assays can be used to identify lymphocyte populations derived from a single cell by detecting the unique V-J gene rearrangements present within these antigen receptor loci.1  This PCR-based assay employs multiple consensus DNA primers that target conserved genetic regions within the  immunoglobulin heavy chain (IGH) gene.

    This master mix can be used to detect and sequence the majority of clonal IGH rearrangements from either genomic DNA (gDNA) or complementary DNA (cDNA).  Clonality is indicated if the master mix generates clonal band(s).  PCR products are differentially labeled with 6FAM and can be analyzed using any platform that has the ability to detect these fluorochromes.  The presence of IGH somatic hypermutation (SHM) is defined as greater or equal to 2% difference from the germline variable (V) gene sequence, whereas less than 2% difference is considered evidence of no somatic hypermutation.2

References

1. Miller, JE, et al. (1999) Molecular Diagnostics. 4(2):101-117.
2. Fais, F., et al. (1998) Journal of Clinical Investigations. 102(8):1515-1525.

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