Lyo-Ready Genotyping Direct DNA qPCR FFPE

PDGFRA1 gene mutations

Lyo-Ready Genotyping Direct qPCR FFPE

KAPA Probe Force qPCR Mix

TaqPath ProAmp Multiplex Master Mix

A/A

G/A

G/G

RFU for Allele 1 - FAM

Healthy FFPE lung tissue (WT) and lung tumour tissue containing the tumor angiogenesis associated PDGFRA1 mutation (Mutant) were digested using MDX168 extraction buffers. 5.2 ng of DNA from digested tissues were in the reaction. Homozygous Allele G ( red ) and allele A ( blue ) and heterozygous Allele G/A ( green ) with a NTC ( black ) and x for undetermined.

ATM gene mutations

T/T

C/T

C/C

RFU for Allele 1 - FAM

Healthy FFPE lung tissue (WT) and lung tumour tissue containing the cancer associated ATM17 mutation (Mutant) were digested using MDX168 extraction buffers. 1.3 ng of DNA from digested tissues were in the reaction. Homozygous Allele C ( red ) and allele T ( blue ) and heterozygous Allele C/T ( green ) with a NTC ( black ) and x for undetermined.

Crudely extracted FFPE DNA vs Purified FFPE DNA – PDGFRA1 Assay

MDX168

KAPA Probe Force qPCR Mix

TaqPath ProAmp Multiplex Master Mix

A/A

G/A

G/G

RFU for Allele 1 - FAM

1.3 ng of DNA from lung tumour FFPE tissue containing the tumoral angiogenesis associated PDGFRA1 mutation (Mutant) was either crudely extracted using the MDX168 Extraction Buffer and analysed with A/ Lyo-Ready Genotyping Direct qPCR FFPE (MDX168) 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 Lyo-Ready Genotyping Direct qPCR FFPE (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.