Instructions for use are available on the product page, downloads section.
This kit contains enough reagents for 24 tests. This means 12 donor/recipient pairs (24 samples).
This kit includes 48 reactions. Our advise is to perform the measurement of both pre and post transplantation samples in triplicate, which means you use 6 reactions per transplant. In total this will allow you to use every marker to test 6 transplants or 6 time points of the same transplant.
The KMRassay® Reference Assay (previously KMRassay® Reference Gene) is a not a gene in itself. It is a marker that will allow the detection of a housekeeping gene.
Therefore the KMRassay® Reference Assay can be used as a control for the presence of any DNA.
The qPCR multiplexes used for genotyping that are contained in the KMRtype kit are not designed, optimized or otherwise validated for use in quantitative analysis of chimeric mixtures. We do not recommend the use of these multiplexes for quantifying mixtures and do not support the use of these reagents for that purpose.
KMRengine is engineered to allow operators to choose with which machine they would like to use for their experiment.
The following instruments are currently compatible with KMRtype and KMRtrack Assays.
- ABI ViiA7 v1.2.4
- ABI 7500 Fast v2.0.6
- ABI 7500 v1.3.1/1.4/2.0.2/2.0.6/2.3
- QuantStudio 5, v1.2
- QuantStudio 6, v1.2/1.3
- QuantStudio 7, v1.2
- QuantStudio 12K, v1.2
- QuantStudio DX, v1.2
- BioRad CFX 96, v1.6/3.0/3.1
For KMRtype you need 4 channels:
- JOE, VIC, HEX or Orange560
- ROX, Texas Red or Red610
- Cy5 or Quasar670 (passive reference)
For KMRtrack you need 2 channels:
- Cy5, Texas red or Quasar670 (passive reference)
They were designed based on population data for European Caucasian, African, American Indian and Japanese populations. While we don’t have clear data for Asian subpopulations, these same set of populations proved well in the Korean population as published in The Journal of Molecular Diagnostics, Vol. 16, No. 6, November 2014 by Seon Young Kim et al. from the Department of Laboratory Medicine, Seoul National University College of Medicine, titled; Chimerism Monitoring after Allogeneic Hematopoietic Stem Cell Transplantation Using Quantitative Real-Time PCR of Biallelic Insertion/Deletion Polymorphisms.
The experiment from DNA to final chimeric mixture make up can be performed in one day. The hands on time will be about 2 hours, 1 hour to set up the genotyping assay and 1 hour to set up the monitoring assay. Analysis is basically instantaneous.
Stock solutions should be stored in containers which minimize light exposure. We have not observed any issues with normal light exposure in routine usage.
In genotyping, 10 ng of input DNA is used per well with the KMRtype reagents. When using KMRtrack reagents, the optimal amount of template DNA to use in chimeric mixture analysis depends on the desired sensitivity detection limit.