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Mutation analyses for BRAF and KRAS were performed for the 28 MSI-H GC patients.
Mutation analyses for MLH1 and MSH2 were performed for the 45 MSI-H GC patients.
For some gRNA transgenic vectors such as CR7T- bam 1, U6B- nos 1 nos 2, and CR7T- nos 1 nos 2, we picked up two independent gRNA transgenic lines for conditional mutation analyses for each transgenic vector.
Mutation analyses for poly(T 11 nucleotide repeats in IVS-4 of MRE11 and poly(A 9 nucleotide repeats in the ORF of RAD50 were performed on genomic DNA extracted using the DNeasy kit (Qiagen, Valencia, CA, USA).
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14 The primer sequences and conditions of PCR for mutation analyses of these genes have been described in previous papers.
We believe that the type of detailed network analyses of gene expression data presented in this paper when combined with next generation sequencing for mutation analyses of individual ovarian adenocarcinoma genomes will help expand our understanding of the origins of ovarian cancer and facilitate the development of more effective therapies.
The 14 POmutationsanalysednalysed for catalytic activity in the present study are derived from gene analyses described in 11 different previously published studies (see Table 1).
We also discuss the importance of a new publicly available and extensively curated database PDmutDB, and the implications of these analyses for mutation analyses in a diagnostic setting.
The authors thank Pauline Chu for immunohistology assistance, Farbod Babrzadeh for assistance with the PIK3CA mutation analyses, and Kyra Heirich for help with manuscript review.
SF and AO were responsible for material support for mutation analyses.
If this study had been prospectively conducted, however, tissue for immunohistochemical and mutation analyses could have been sampled for all tumours, thereby assuring validity of diagnoses of both patient groups studied.
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