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CAT #: 21010180

Hypermutation Mix 2 v2.0 - Unlabeled for Gel Detection

Product Use

Hypermutation Mix 2 v2.0 – Unlabeled for Gel 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 can be analyzed using standard gel electrophoresis using heteroduplex analysis with ethidium staining.  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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