Application Guide - QIAxcel Advanced

48 QIAxcel Advanced Application Guide 10/2016 Ratio charts Coffalyser ratio charts are an additional overview of MLPA results and were available for the results from the sequencer only (Figure 4). Complementary to electropherograms, the ratio charts re-interpret the same exonic information. The displayed confidence interval enables assessment of the success of the MLPA run. Mutation state is indicated by the ratio. Ratios of around 0.5 suggest a heterozygous deletion of a specific allele (Figure 4B, Exons 2–18). Duplication of specific alleles are shown with a ratio of around 1.5 (Figure 4C, Exons 11–13). APC duplication Q-64 (DNA) 97 bp 93 bp 158 bp 163 bp 135 bp 197 bp 178 bp 229 bp 246 bp 254 bp 251 bp 264 bp 270 bp 274 bp 289 bp 311 bp 297 bp 308 bp 319 bp 331 bp 338 bp 344 bp 354 bp 372 bp 379 bp 384 bp 418 bp 421 bp 400 bp 426 bp 432 bp 449 bp Q-70 (DNA) Q-82 (DNA) CARM1-1 (DD) JPH3-up (DD) IL1B-4 (Lig) MUTYH-9 APC-2 APC-2 APC-3 APC-4 APC-5 APC-6 APC-7 APC-8 APC-9 APC-1 APC-1 APC-10 APC-11 APC-12 APC-13 APC-14 APC-15 APC-16 APC-18 APC-18 APC-18 APC-18 (WT) APC-18 (WT) APC-intr.13 APC-17 MUTYH-1 MUTYH-4 AMOT-4 (XC) Reference* Reference* Reference* Reference* Reference* Reference* Reference* Reference* Reference* Reference* Q-80 (DNA) Relative fluorescence units 64 70 82 80 88 130 136 142 150 157 171 178 186 193 202 211 220 229 238 216 253 260 267 274 282 289 297 306 312 321 328 337 346 356 365 373 382 391 400 409 418 427 445 92 96 100 A Relative fluorescence units (RFU x 1EO) B 0.130 0.120 0.110 0.100 0.090 0.080 0.070 0.060 0.050 0.040 0.030 0.020 0.010 0.000 5500 5000 4500 4000 3500 3000 2500 2000 1500 1000 5000 0 APC duplication Q-64 (DNA) 97 bp 93 bp 158 bp 163 bp 135 bp 197 bp 178 bp 229 bp 246 bp 254 bp 251 bp 264 bp 270 bp 274 bp 289 bp 311 bp 297 bp 308 bp 319 bp 331 bp 338 bp 344 bp 354 bp 372 bp 379 bp 384 bp 418 bp 421 bp 400 bp 426 bp 432 bp 449 bp Q-70 (DNA) Q-82 (DNA) CARM1-1 (DD) JPH3-up (DD) IL1B-4 (Lig) MUTYH-9 APC-2 APC-2 APC-3 APC-4 APC-5 APC-6 APC-7 APC-8 APC-9 APC-1 APC-1 APC-10 APC-11 APC-12 APC-13 APC-14 APC-15 APC-16 APC-18 APC-18 APC-18 APC-18 (WT) APC-18 (WT) APC-intr.13 APC-17 MUTYH-1 MUTYH-4 AMOT-4 (XC) Reference* Reference* Reference* Reference* Reference* Reference* Reference* Reference* Reference* Reference* Q-80 (DNA) Relative fluorescence units 64 70 82 80 88 130 136 142 150 157 171 178 186 193 202 211 220 229 238 216 253 260 267 274 282 289 297 306 312 321 328 337 346 356 365 373 382 391 400 409 418 427 445 92 96 100 A Relative fluorescence units (RFU x 1EO) B 0.130 0.120 0.110 0.100 0.090 0.080 0.070 0.060 0.050 0.040 0.030 0.020 0.010 0.000 5500 5000 4500 4000 3500 3000 2500 2000 1500 1000 5000 0 Figure 3. Comparable pattern of peaks for APC duplication using ABI PRISM 3100 Genetic Analyzer and QIAxcel Advanced System. A Electropherogram from Coffalyser software analysis. B Electropherogram from the QIAxcel ScreenGel 1.2 software analysis. Green circles show reference peaks. Black arrows indicate APC peaks increased compared with the corresponding APC control electropherograms in Figure 1. Arrows are similarly placed between electropherograms A and B.

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