High Sensitivity and Reproducibility in Assays Detecting High or Low copies of DNA Templates in 15 mg/mL Human Stool
The activity of Lyo-Ready ™ Genotyping Direct qPCR Stool (blue) was tested in a qPCR assay using a 10-fold serial dilution of synthetic DNA (10,000, 1000, 100, 10 and 1 copies respectively), in the presence of 15 mg/mL human stool, and compared to KAPA Probe Force qPCR Mix ( green ), TaqPath ™ ProAmp ™ Multiplex Master Mix ( black ) and Type-it Fast SNP Probe PCR ( turquoise ). The results illustrate that Lyo-Ready Genotyping Direct qPCR Stool can sensitively and reproducibly detect high or low copies of DNA templates in the presence of PCR inhibitors.
Stable Shelf Life for up to 18 months After Lyophilization
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B)
Lyo-Ready ™ Genotyping Direct qPCR Stool was lyophilized, and the stability of the dried assays ( red ) was tested after 1 month at 37°C against fresh liquid mix ( blue ) with A) purified DNA target, and B) in the presence of 10 mg/mL human stool. The lyophilized mixes retain the same performance as the liquid mix, and the same accurate allelic discrimination where mutant allele-specific amplification is retained while WT non-specific amplification is not observed. Results suggest that Lyo-Ready ™ Genotyping Direct qPCR Stool mix has a projected stability of more than 18 months at ambient temperature.
1. N CCN clinical cancer practice guideline in oncology (NCCN Guidelines) Ver.4-June 15, 2020 http://nccn.org/professionals/physician_gls/pdf/colon.pdf 2. Kim, J.G., et al. TP53 codon 72 polymorphism associated with prognosis in patients with advanced gastric cancer treated with paclitaxel and cisplatin. Cancer Chemother Pharmacol . 2009 Jul;64(2):355-60. doi: 10.1007/s00280-008-0879-3 3. Shen, J., et al. Formation of A2143G mutation of 23S rRNA in progression of clarithromycin-resistance in Helicobacter pylori 26695. Microb Drug Resist . 2005, 11: 100-106. 10.1089/mdr.2005.11.100. 4. Cebula, T.A., et al. Simultaneous Identification of Strains of Escherichia coli Serotype O157:H7 and Their Shiga-Like Toxin Type by Mismatch Amplification Mutation Assay-Multiplex PCR. J. Clin. Microbiol. , Jan. 1995, p. 248–250
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