1. A B C D Reference Standard (HD728)
No dilution
Dilution 1:2
Dilution 1:4 D N°
Gene
Variant
Theoric AF
Measured AF
No dilution
Dilution 1:2
Dilution 1:4 1
NRAS
p.Q61K
5.0%
4.4%
2.4%
0.9% 2
*DDR2
p.M117I NR
25.3%
13.7%
6.1% 3
KRAS
p.G13D
25.0%
22.5%
13.6%
6.2% 4
p.G12A
5.3%
2.9%
1.2% 5
p.G12R
5.2%
2.2%
1.1% 6
*MAP2K1
p.Q56P
18.8%
10.6%
4.2% 7
*TP53
p.? c.376-7C>T
19.2%
10.9%
5.4% 8
PI3KCA
p.H1047R
30.0%
32.4%
17.7%
8.1% 9
*CTNNB1
p.S33Y
19.8%
8.6%
4.5% 10
BRAF
p.V600E
8.0%
9.9%
5.6%
4.0% 11
p.V600K
3.1%
1.9%
1.7% 12
EGFR
p.G719S
16.7%
19.4%
9.6% 13
p.T790M
4.1%
2.3%
Other variants ND
Supplemental Data Figure 7: Dilution of the reference standards (HD728). Result plots of HD728 dilutions (A–C). Numbers in the plots identify different mutations detailed in panel D. Red dots indicate a hotspot mutation; blue dots indicate a non-hotspot mutation. Panel D shows the list of detected mutations, the measured allele frequencies, and the theoretical allele frequencies given by the manufacturer. *Indicates a mutation found in the exome sequencing performed by the manufacturer, but with a non-reported (NR) allelic frequency.