Implementation of a robotized real-time PCR setup for the use of the Quantifiler™ Human DNA Quantification Kit

Evaluation of droplet digital PCR and next generation sequencing for characterizing DNA reference material for KRAS mutation detection

“…To assess the KRAS copy number variation in each of the 7 tumor cell lines, a duplex ddPCR assay targeting KRAS mutant and wild type was used to report the total KRAS copy number concentration. In addition, a Quantifiler Human DNA Quantification kit (Thermo Scientific) was used on the QX100 to quantify a single copy target of the human telomerase reverse transcriptase gene ( hTERT ) 26 which can serve as a reference single copy gene. The ratio of KRAS to hTERT , which clarifies the KRAS copy number variation (Table 1 ), was very close to 1 for cell line RPMI-8226, SNU-C2B, SW1573, A549, HCT116 and 293 T, indicating that no variation in KRAS copy number occurs.…”

Evaluation of a semi-automated, magnetic bead-based DNA extraction method for genetic fingerprinting of forensic casework samples

“…Cross-contamination tests were carried out by DNA extractions of positive and negative control samples arranged in checked pattern. DNA-quantification was determined by real-time PCR using the AB Quantifiler TM Human DNA Quantification Kit [1] (Applied Biosystems). For differential extraction of mixed sperm/vaginal secretion stains, samples were incubated 10 min at 56 8C and isolated from the trace carrier.…”

Reliability and Cost of GMO Detection

“…Automation is a promising approach to achieve a cost reduction in conjunction with equivalent or even increased reliability compared to manual handling. While automated DNA extraction and PCR setup are already well established and common practice in other fi elds of PCR diagnostics (CRL -GMFF, 2008 ;Greenspoon et al, 2006 ;Schwenzer et al, 2008 ;Smith et al, 2004 ;Wang and Ginocchio, 2009 ), automation of GMO detection is still in its early stages. Within the Co -Extra project, the feasibility, economic aspects and implications of automation in GMO analysis by automation of DNA extraction and PCR setup were investigated, and experimental details are available in the deliverable D5.12 (Gruden et al, 2009b ).…”