Table 1.
HRM | Pyrosequencing | |||||||
---|---|---|---|---|---|---|---|---|
Run 1 | Run 2 | |||||||
Sample | cDNA [ng/μl] | Run 1 | Run 2 | Result | % mut. Alleles | Result | % mut. Alleles | Final result |
1 | 11 | failed | WT a | WT | WT | |||
2 | 31 | WT a | WT | WT | ||||
3 | 10 | WT a | G12C | 13.4 | mut | |||
4 | 93 | WT a | WT | WT | ||||
5 | 26 | WT a | WT | WT | ||||
6 | 27 | WT a | WT | WT | ||||
7 | 10 | WT a | WT | WT | ||||
8 | 43 | mut | G13D | 73.9 | mut | |||
9 | 85 | failed | WT | WT | WT | |||
10 | N/A | mut | G13D | 44.3 | mut | |||
11 | N/A | failed | G12V | 2.6 | WT b | |||
12 | N/A | WT | WT | WT | ||||
13 | N/A | mut | G12V | 41.9 | mut | |||
14 | 126 | mut | failed | G12D | 65.7 | mut | ||
15 | 133 | mut | G12D | 74.1 | mut | |||
16 | 97 | mut | G13D | 52.8 | mut | |||
17 | 47 | failed | WT | WT | WT | |||
18 | 14 | failed | WT | WT | WT | |||
19 | 44 | WT a | WT | WT | ||||
20 | 20 | failed | WT a | WT | WT | |||
21 | N/A | failed | WT | WT | ||||
22 | 138 | mut | G12V | 12.9 | mut | |||
23 | 325 | mut | G12D | 56.1 | mut | |||
24 | 140 | WT a | WT | WT | ||||
25 | 7 | mut | G12C | 61.2 | mut | |||
26 | 4 | WT a | WT | WT | ||||
27 | 27 | WT a | WT | WT | ||||
28 | 13 | WT a | WT | WT | ||||
29 | 207 | mut | G12D | 54.5 | mut | |||
30 | 10 | failed | WT | G12V | 1.5 | WT b | ||
31 | 54 | WT a | WT | WT | ||||
32 | 120 | mut | G12V | 29.2 | mut | |||
33 | 33 | failed | mut | G12C | 33.5 | mut | ||
34 | N/A | mut | G12C | 71.4 | mut | |||
35 | N/A | WT | WT | WT | ||||
36 | N/A | failed | WT | WT | ||||
37 | 67 | WT a | WT | WT | ||||
38 | 137 | mut | G12C | 76.7 | mut | |||
39 | 63 | mut | G12A | 56 | mut | |||
40 | 13 | WT a | WT | WT | ||||
41 | 113 | WT a | WT | WT | ||||
42 | 82 | mut | G12C | 75.4 | mut | |||
43 | 39 | WT | WT | WT | ||||
44 | 7 | WT a | G12S | 2 | WT b | |||
45 | 23 | WT a | WT | WT | ||||
46 | 24 | faileda | G13D | 3.5 | WT b | |||
47 | 9 | WT a | WT | WT | ||||
48 | 34 | WT a | G12S | 2 | WT b | |||
49 | 25 | WT a | WT | WT | ||||
50 | 17 | WT | WT | WT | ||||
51 | 5 | WT a | WT | WT | ||||
52 | 29 | WT a | WT | WT | ||||
53 | 31 | mut | G12D | 71.3 | mut | |||
54 | 68 | WT | WT | WT | ||||
55 | 93 | mut | G12D | 74.2 | mut | |||
56 | 221 | WT | WT | WT | ||||
57 | 83 | WT | WT | WT | ||||
58 | N/A | WT a | G12V | 1.2 | WT b | |||
59 | 35 | mut | G12D | 83.3 | mut | |||
60 | N/A | WT | WT | WT | ||||
61 | 37 | WT a | WT | WT |
aSkewed HRM curve
bLOD/threshold for potential low level mutation (cf. Therascreen KRAS Pyro Kit handbook version 1, July 2011): G12D 2.2 %/5.2 %, G12V 1.0 %/4.0 %, G12C 2.1 %/5.1 %, G12S 1.9 %/4.9 %, G13D 1.9 %/4.9 %