Non-B Genomic Detection Kit

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Available in classical PCR or qPCR format

ACCUMOL's Non-B Genomic Detection Kit allows the detection of Non-B Cell DNA in genomic DNA samples prepared from sorted B Cells. During normal B Cell differentiation, rearrangement of the IGH genes results in the excision of genomic DNA. This modification of the genome takes place on both alleles. The Non-B Genomic Detection Kit is designed to amplify a sequence located in the excised region, that is no longer part of the genome of mature B Cells. The detection of the target sequence in the gDNA sample indicates that contaminating Non-B Cells were present in the original B Cell preparation.

Since the test is performed on DNA, ACCUMOL's Non-B Genomic Detection Kit is not affected by the presence of beads or antibodies on the surface of the sorted cells, and can be performed regardless of the sorting method used (flow cytometry, magnetic beads, density-based, adhesion, etc...). And because ACCUMOL's Non-B Genomic Detection Kit detects the purity information directly at the level of the DNA sample, the sample can be analyzed at any point in time, even years after the actual cell sorting took place.

Classical PCR format

The Non-B Genomic Detection Kit is provided ready to use as a reaction mix aliquoted in 0.2 mL PCR tubes. Just add the template (about 2 ng of gDNA), and load the reaction tube in your thermocycler. The PCR program we recommend is a standard 30 cycles 3-step program. The Non-B Genomic Detection Kit is very robust and will perform under a wide range of PCR conditions.

Note that this kit has been primarily designed to integrate seamlessly in chimerism analysis workflows. PCR programs used in most chimerism analysis protocols are compatible with the Non-B Genomic Detection Kit. If you are performing chimerism analysis, you will likely not need to re-program your thermocycler. The reaction can be run along with your chimerism samples, with your existing settings.

PCR products analysis is performed on a sequencer or genetic analyser, using conditions similar to the ones used in most chimerism analysis procedures.

Catalog numbers: 010022 (96 reactions), 010023 (32 reactions)

qPCR format

The qPCR Non-B Genomic Detection Kit is provided as a reaction mix ready to be dispensed in qPCR tubes or plates. Just add the template (about 2 ng of gDNA), and load the reaction tubes or plates in your qPCR instrument. Following amplification, the products are analyzed by melting curve analysis. This kit is compatible with most qPCR instruments, using standard or FAST programs.

Catalog numbers:
No Passive Reference: 030122 (96 reactions), 030123 (32 reactions)
Low Passive Reference: 030132 (96 reactions), 030133 (32 reactions)
Passive Reference: 030142 (96 reactions), 030143 (32 reactions)

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Benefits

  • Assess the purity of sorted B cells directly in the genomic DNA sample.
  • Save cells; stop performing flow cytometry
  • Works with as little as 2 ng of DNA (only 1,000 cells!)
  • Just add sample to ready-to-use mix
  • Use the equipment already in your lab
  • Easy integration within existing workflow
  • Can be performed years after cell sorting

Application