Table 4 Effect of GeneClean purification and tumour heterogeneity on Cast-PCR and Sanger sequencing sensitivity.

Case	SANGER		Cast-PCR
Case	GC-purified	Not purified	GC-purified	ΔCT	Not purified	ΔCT
2	V600E	Failed	V600E	1.95	V600E	1.9
3 sample A	V600E	Failed	V600E	1.85	V600E	2.53
3 sample B	V600E	Failed	V600E	2.22	V600E	2.32
4	V600E	Failed	V600E	1.56	V600E	1.46
6	uncertain	Not mutated	V600E	2.01	V600E	6.92
7	Not mutated	Failed	V600E	5.17	Not mutated
8 sample A	V600E	V600E	V600E	1.74	V600E	2.09
8 sample B	V600E	V600E	V600E	2.06	V600E	1.56
9 sample A	Not mutated	Failed	V600E	5.21	Failed
9 sample B	Uncertain	Failed	V600E	4.40	Failed
9 sample C	V600E	Failed	V600E	3.44	Failed
9 sample D	V600E	Failed	V600E	3.33	Failed
10 sample A	V600E	V600E	V600E	1.58	V600E	2.30
10 sample B	V600E	V600E	V600E	0.10	V600E	1.32
11 primary	Not mutated	Failed	V600E	4.69	V600E	5.00
11 lymphnode	Not mutated	Failed	V600E	2.72	Not done
15	V600E	Failed	V600E	3.33	V600E	5.03
22	uncertain	Failed	V600K	6.93	V600K	8.3
23	V600K	V600K	V600K	2.4	V600K	3.22
24 primary	V600K	Not done	V600K	0.77	Not done
24 lymphnode	Not mutated	Not done	Not mutated		Not done
25	Failed	Failed	V600K	2.04	V600K	6.03
29	V600K	V600K	V600K	3.77	V600K	4.05
31 sample A	Not mutated	Failed	Not mutated		Not mutated
31 sample B	Not mutated	Not done	Not mutated		Not done
32	Not mutated	Failed	Not mutated		Not mutated
38	Not mutated	Failed	Not mutated		Not mutated
39	Not mutated	Failed	Not mutated		Not mutated
40	Not mutated	Failed	Not mutated		Not mutated
41 sample A	Not mutated	Failed	Not mutated		Not mutated
41 sample B	Not mutated	Not done	Not mutated		Not done
41 sample C	Not mutated	Not done	Not mutated		Not done
42	Not mutated	Not mutated	Not mutated		Not mutated
43 block 1	Not mutated	Failed	Not mutated		Not mutated
43 block 2	Not mutated	Not done	Not mutated		Not done

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