Download Supplemental Figure 2

K-RAS exon 2
EGFR exon 19
conventional-PCR
conventional-PCR
≅ 5% C>T
≅ 5% C>A
≅ 44% C>T
≅ 35% C>A
TP53 exon 5
conventional-PCR
≅ 5% G>A
≅ 5% G>T
antisense
TT-fast-COLD-PCR
TT-fast-COLD-PCR
≅ 58% C>T
TP53 exon 6
TP53 exon 7
conventional-PCR
conventional-PCR
≅ 5% G>T
conventional-PCR
≅ 5% C>T
antisense
TT-fast-COLD-PCR
EGFR exon 20
TT-fast-COLD-PCR
≅ 67% G>A
≅ 78% G>T
TP53 exon 8
TP53 exon 9
conventional-PCR
conventional-PCR
≅ 5% C>T
≅ 5% C>T
≅ 5% C>T
antisense
TT-fast-COLD-PCR
≅ 63% G>T
TT-fast-COLD-PCR
≅ 33% C>T
TT-fast-COLD-PCR
≅79% C>T
TT-fast-COLD-PCR
≅82% C>T
Supplemental Figure 2 (expanded version of Figure 2). Sanger sequencing following 50-plex fast-TT-COLD-PCR or, alternatively, conventional 50-plex PCR. A mixture
of genomic DNA from several cell lines (~5% mutation abundance) was pre-amplified using a 50-plex PCR using gene specific primers containing common sequences on the 5’
end. The sample was then split in eight parallel tubes and fast-TT-COLD-PCR was then performed using a single primer approach to co-amplify all 50 amplicons. Eight mutated
sequences within the 50 amplicons were evaluated for. These targets represent sequences for which mutation-containing DNA and appropriate primers were available.