Use of the QIAGEN GeneReader NGS system for detection of KRAS mutations, validated by the QIAGEN Therascreen PCR kit and alternative NGS platform

Table 1 Parameter and sequencing coverage of Actionable Insight Tumor Panel

Parameter	Details
Panel size	12 genes/16.7 kb
Insight size	773 unique variant positions
Amplicons	330
Variant allele fraction detection limit	5%
Frequency cut-off and amplicon coverage	>500×: 96.4% (A), 92.0% (B)
Frequency cut-off and amplicon coverage	>200×: 98.5% (A), 97.2% (B)
Frequency cut-off and variant insight coverage	>500×: 99.8% (A), 98.6% (B)
Frequency cut-off and variant insight coverage	>200×: 99.9% (A), 99.8% (B)

Positive samples included into the study have all been confirmed with Sanger sequencing and passed QuantiMIZE (<0.4). (A) An average of 12 NA12878 samples, (B) average of 43 colorectal cancers FFPE samples (ages 3-20 years)

ISSN: 1471-2407