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In our research [12], we precisely described the mutations performed by each operator.
The curves are based on EPSA correlation scores for each of the five oncogenic mutations performed by Bild et al. (see Methods).
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The initial diagnostic testing for BRAF mutations was performed by primer extension reaction (BRAF mutation analysis reagents, Applied Biosystems, CA, USA).
Because the initial screening for BRCA1 and BRCA2 mutations was performed by local diagnostic laboratories using protocols of varying sensitivity, a systematic attempt is underway to identify additional mutations using full sequencing of BRCA1 and BRCA2, which has been performed in 168 cases (from 163 families).
Mutation analyses of exons 2 11 of the TP53 gene in the 30 cases with BRCA1 germ line mutations were performed by TTGE followed by sequencing.
Most testing for JAK2 mutations is performed by analyzing the genomic DNA of the JAK2 gene.
Ser27Ala, Ser47Ala and Thr530Ala mutations were performed by using site-directed mutagenesis.
Mapping and genotyping of the new mutations were performed by using a total of 174 DNA samples classified as indicated above.
Classification of BRCA1/2 mutations was performed by SH and OMS.
Screening for mutations was performed by high-resolution melting curve analysis on a Lightscanner (Idaho Technology , Inc.
The screening of VHL mutations was performed by single-strand conformational polymorphism analysis (SSCP).
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