FAP MASTR Research Application – For detection of germline variants and SNVs in APC and MUTYH in blood-derived DNA. FAP MASTR is a disease research panel for the identification of variants in the complete coding regions of the APC and MUTYH genes, which are associated with familial adenomatous polyposis (FAP). This panel is ready to use and offers robust performance with minimum hands-on time. It consists of all the reagents necessary to enable multiplex amplification of 52 amplicons for complete exon coverage of the APC and MUTYH genes. Assay Characteristics Genes analyzed Genomic region analyzed Number of amplicons Amplicon length Number of plexes Designed to be compatible with full coding region of APC and MUTYH 14.7 kb 52 245-455 bp 3 Illumina MiSeq Performance Uniformity of amplification (0.2x mean coverage) 99.7 % On target read count > 96 % DNA input Number of samples/run (20 reads/allele): *only 192 MID combinations available. as low as 20 ng per plex reaction Illumina MiSeq V2: 1,200* Illumina MiSeq V3: 2,200* Performance cont. 14% Percentage of amplicons Graph presenting the read counts for all FAP MASTR amplicons, showing their outstanding uniform representation. To allow comparison between samples, the read counts were normalized to a coverage of 100. The data is based on 22 samples collected from three centers. *3’ UTR of EPCAM only 12% 10% 8% 6% 4% 2% Read counts MASTR Workflow Multiplex PCR Plan run Multiplex PCR Dilution Quality Control Mixing Clean up Universal PCR Universal PCR Quality Control Pooling NGS SEQUENCING Mixing plexes of single sample Purification Ordering Information Cat. No. Product Name Reactions MR-0040.008 FAP MASTR 3 • MID (Molecular Identifiers) kits are necessary to complete the workflow www.agilent.com Not for EU [email protected] For Research Use Only. Not for use in diagnostic procedures. This information is subject to change without notice. PR7000-0725 © Agilent Technologies, Inc. 2017 Published in the USA, September, 2017 5991-8362ENUC Equimolar pooling DATA ANALYSIS 400 380 360 340 320 300 280 260 240 220 200 180 160 140 120 100 80 60 40 0 20 0%