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. 2010 May 22;101(9):2005–2010. doi: 10.1111/j.1349-7006.2010.01627.x

Figure 2.

Figure 2

 Correlation of mutation detection using mutation‐specific PCR and pyrosequencing. (a) Sensitivity of assay for mutation detection by pyrosequencing for T315I was established by dilution study on involved patient sample and compared to expected dilution curve. RNA from a patient sample with predominantly mutated transcript (approximately 75%) was diluted into HL‐60 RNA. (b) Correlation of relative mutation levels in quantitative (q)RT‐PCR and pyrosequencing assays was tested in 14 patient samples. Dotted lines indicate 95% confidence intervals. (c) Sensitivity of assay for mutation detection by pyrosequencing for G250E was established by dilution study on involved patient sample and compared to expected dilution curve. RNA from a patient sample demonstrating only mutated G250E transcript was diluted into HL‐60 RNA. (d) Correlation of relative mutation levels in qRT‐PCR and pyrosequencing assays was tested in 18 patient samples. (e) Precision of the T315I pyrosequencing assay quantitation was assessed in 98 PCR reactions split at the cDNA step and separately amplified in first‐ and second‐round PCR and pyrosequenced.