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The number of mutations was recorded, and a gene was considered lethal if there were more than 10 mutations in it.
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Although we have not performed sequencing on matched normal blood samples in this study, the mutations identified were assumed to be somatic because they are located in the common mutational hotspot observed in uterine leiomyomas and fibroadenomas, and none of the mutations were recorded as germline variants in the databases.
Mutations were recorded according to the International AIDS Society-USA consensus [23].
Some of these mutations are recorded in MITOMAP (http://www.mitomap.org/MITOMAP) as being related to some diseases, although none of them are labelled as "confirmed" pathogenic mutations.
The dystrophin gene is one of the largest in the human genome (approximately 2.5 million base pairs, encoding 79 exons) and many thousands of mutations are recorded [7].
In 2010, single mutations were recorded in some isolates in addition to the presence of the HP cleavage site pattern of 2006 in all years which means that different genotypes of H5N1 virus are co-circulating in different areas.
In 2009, aa changes were more frequent in the cleavage region of the HA and more than one aa mutations were recorded in one isolate compared to the pattern of 2006 which represent the HPAIV ideal pattern.
Also, while initially, the most commonly occurring mutations were recorded; now there is potentially a bias toward only novel mutations being recorded in the Leiden database.
Losses of mutation classes and fixations of mutations were recorded separately.
Known somatic mutations were recorded for 422 distinct genes which were identified by NCBI Gene ID.
Positions, types, and associated Phred scores of all mutations were recorded.
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