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We calculated that microcolony-specific increases in mutation frequency were at least 100-fold compared with planktonically grown cultures.
Therefore microcolony-specific increases in mutation frequency were at least 100-fold in our experiments, and may be up to 1800 fold higher than that observed in planktonic culture.
Moreover, in our experiments, we used different polymerases with different error rate (see M/M) and no significant differences in mutation frequency were observed, arguing against the generation of the mutations during PCR amplification.
Profound differences in mutation frequency were observed between various histopathological subtypes of tumors.
Categorical data (for example, sex, ethnicity, stage, mutation frequency) were compared using a chi-squared test.
Differences in forward mutation frequency were observed between the two different batches of MMC and not between the optimal dose and half the optimal dose (375 µM).
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The U2AF1 mutation frequency was higher, with different frequencies in the mutated sites of U2AF1 (S34Y, 6/25; S34F, 11/25; and Q157P 8/25).
The HPRT mutation frequency was evaluated as described (van der Kuip et al., 2004).
Moreover, this mutation frequency was dependent on the ability of Cas9 to introduce a dsDNA break at the target site.
As shown in Figure 1c, HPRT mutation frequency was consistently elevated in cells lacking c-Abl kinase activity.
As expected, in cells expressing imatinib-resistant Bcr-Abl and imatinib-sensitive c-Abl, the HPRT mutation frequency is significantly enhanced in the presence of imatinib (Figure 1b).
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