Lyo-Ready Genotyping Direct qPCR FFPE, 4x

Reliable genotyping directly from crude FFPE samples

Product Description

To minimize the workflow to isolate DNA from FFPE tissue, Meridian has created the Lyo-Ready Genotyping Direct qPCR FFPE, this allows digestion of the tissue in a single tube, without the need for purification, greatly reducing the risk of sample damage, loss and contamination whilst simplifying and accelerating the analysis process. It can also be used in a liquid format or lyophilized format to create ambient-temperature stable assays, making it ideal for high throughput.

For more details please see user manual above and the Lyophilization & Post-Lyophilization User Guideline.

Product Highlights

• Ultra-sensitive detection down to less than 1 copy
• Suitable for crude FFPE samples
• Tight fluorescence clusters with clear allele discrimination, perfect for difficult SNPs
• Can be used as a liquid or lyophilized, ideal for high-throughput testing
  
  

SNPs (single nucleotide polymorphisms) or point mutations are the most common types of genetic variation and comprise the major part of phenotype diversity between individuals. SNPs can be responsible for both resistance or susceptibility to a certain disease and once individual SNPs have been identified by sequencing, they can be used for pharmacogenetics, in evaluating and predicting a patient’s response to treatment and risk of adverse events, or for diagnostics, such as in cystic fibrosis screening. Although blood is an ideal source for genotyping analysis, formalin-fixed paraffin-embedded (FFPE) tumor tissues are also widely used and constitute valuable resources for retrospective studies due to the possibility of long-term storage at room temperature. Despite the extremely time-consuming and labour-intensive workflows, success rate of genotyping assays using DNA purified from such samples remains low.

Supporting Data

Fig.1. Genotyping of FFPE lung tissue samples. Healthy FFPE lung tissue (WT) and lung tumor tissue containing the tumor angiogenesis associated PDGFRA1 mutation were run with A/ Lyo-Ready Genotyping Direct qPCR FFPE (MDX168) following extraction with the extraction buffers provided or purified using Qiagen QIAamp DNA FFPE Tissue Extraction Kit and analysed with B/ KAPA Probe Force qPCR Mix (Roche) and C/ TaqPath ProAmp Multiplex Master Mix (ThermoFisher). Homozygous Allele G (red) and allele A (blue) and heterozygous Allele G/A (green) with a NTC (black) and x for undetermined. The results illustrate that MDX168 shows greater allelic discrimination and tighter clustering even in the presence of crude FFPE tissue extracts in comparison to other commercially available master mixes using purified gDNA.