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The improvement in MPS platforms and the forthcoming new technologies constantly increase the length of the reads, and thus the computational tool provided in this paper when applied to such longer reads will give a more comprehensive view regarding co-occurrence of several mutations. In our case, a very strong correlation was observed between mutations altering nucleotides 1308 and 1313 (Table?3). In September 2009, these two mutations, C1308T and T1313C, appeared almost exclusively together. Each one of the two amino acids encoded by the http://www.selleck.cn/products/ON-01910.html altered nucleotide triplets has direct and close interactions with various TKIs (Fig.?4F). The two mutations may act in a cooperative way in order to effectively alter the binding affinity with the inhibitors. Utilizing this double mutant also allows us to draw conclusions regarding the emergence of mutated clones prior to treatment. While DCMD successfully http://www.selleckchem.com/products/loxo-101.html considers sequencing-induced errors, we are still limited by the error rate introduced by reverse transcription and PCR amplification, and we are therefore obliged to pick a conservative mutation rate threshold to eliminate false positives. Under this threshold, we could not detect mutations at time of diagnosis. In order to overcome this limitation, we considered co-existence of two mutations, specifically the C1308T and T1313C double mutant, in the pre-treatment sample. By assuming independence in the distribution of sequencing errors in two positions, the number of double http://www.selleckchem.com/products/abc294640.html mutated reads significantly exceeds the number expected by chance (P?