Performance Data
Lyo-Ready Genotyping Direct qPCR Urine combines the latest advances in buffer chemistry and PCR enhancers, together with an optimized antibody-mediated hot-start polymerase, for fast, precise, and highly reproducible allelic discrimination and cluster separation with SNP detection assays, even in the presence of PCR inhibitors found in urine (and transport media).
Detection of Human Papillomavirus (HPV) Type 16 from 40% Universal Transport Media (UTM)
A)
B)
Detection of human papillomavirus (HPV) type 16 E6-T350G mutation was performed using A) Lyo-Ready ™ Genotyping Direct qPCR Urine (MDX158), B/ KAPA Probe Force qPCR Mix (Roche), C) TaqPath ™ ProAmp ™ Multiplex Master Mix (ThermoFisher) and D) Type-it Fast SNP Probe PCR Kits (Qiagen) in presence of 40% Universal Transport Media (UTM). Homozygous samples for Allele G ( blue ) and Allele T ( red ) and heterozygous samples for Allele G/T ( green ) were compared with a NTC ( black ) and x for undetermined. The presence of the 16 E6-T350G change may have biological advantages that promote cell immortalization in cervical and oropharyngeal cancer 1 . The results illustrate ability of Lyo-Ready ™ Genotyping Direct qPCR Urine to form tight clustering and so accurate allelic discrimination in the presence of clinically relevant mutations directly from universal transport media.
C)
D)
Allele G | Allele T | Allele G/T | NTC
Detection of Pancreatic Cancer-associated KRAS-G12D Mutation from 10% Pooled Human Urine
A)
B)
Detection of pancreatic cancer-associated KRAS-G12D mutation (SNP accession number: rs121913529) from DNA of pancreatic cancer cell line SU.86.86 was performed using A/ Lyo-Ready ™ Genotyping Direct qPCR Urine, B/ Kapa Probe Force, C/ TaqPath ™ and D/ Type-it Fast Kits, in presence of 10% pooled human urine. Homozygous Allele A ( blue ) and Allele G ( red ) and heterozygous Allele A/G ( green ) with a NTC ( black ) and x for undetermined. The KRAS-G12D mutation is present in approximately 35% of people diagnosed with pancreatic cancer, however new studies in mice have identified a promising experimental drug that directly targets pancreatic tumors with this mutation 2 . The results illustrate the ability of Lyo-Ready ™ Genotyping Direct qPCR Urine to detect and accurately discriminate cancer-associated SNPs from human cancer cells directly from crude human urine samples.
C)
D)
Allele A | Allele G | Allele A/G | NTC
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