Quantitative Molecular Reference Standards

Do You Monitor the Limit of Detection of Your Molecular Assays?
Verify the true sensitivity of your molecular standards every day

Analytical Validation & Monitoring

A wide range of platforms is currently in use for genotyping, including Sanger sequencing, q-PCR, pyrosequencing, and more recently,  next-generation sequencing technologies such as Life Technologies’ IonTorrent and Illumina's MiSeq and HiSeq platforms

A major challenge faced by scientists is that each of these platforms has differing sensitivities, and limits of detection, and this, in conjunction with variation in sample preparation, DNA yield and concentration, can limit the ability to accurately detect low frequency mutations. It is therefore essential to validate your chosen platform using high quality reference material.

Horizon Diagnostics has developed a range of genomic DNA reference standards specifically designed for the validation of genotyping assays and for comparing between different platforms. 

Horizon Diagnostics’ Reference Standards are high quality genomic DNA aliquots extracted directly from genetically engineered isogenic human cell lines. The cell lines are generated by engineering the mutation of interest (e.g. BRAF V600E) into a human cell line at the appropriate gene loci using the GENESIS™ gene engineering platform. This process introduces the desired mutation into the cell’s genome exactly as it would occur in a real tumor, yielding one ‘mutant’ cell line and one matched ‘normal’ or parental cell line, differing only by the specific alteration precisely introduced into the genome.

The engineered mutant DNA is then blended with DNA from the normal cell line to achieve the desired ratio between the two alleles, ranging from 50% mutant down to 1%. HDx precisely validates these reference materials using multi-parametric analysis, including Droplet Digital PCR, to confirm allele abundance with a precision of 0.1%.

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