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In this work, a TILLING population for hexaploid oat, consisting of 2550 different mutagenised seed lines was developed and the mutation frequency determined.
The mutation frequency determined in this investigation was considerably lower than reported in the general literature; nevertheless, two other studies documented comparably lower p53 mutation frequencies of 8 and 12% (also at codon 245) (Casson et al, 1991; Gonzalez et al, 1997).
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Because the error-prone polymerase UmuC plays a key role in accelerating the mutation frequency, we determined the influence of phototreatment on the development of spontaneous resistance to rifampicin.
Then, the mutation frequency was determined as the ratio of the number of rifampin resistant cells and the number of viable cells.
Mutation frequency is determined by fluctuation assays.
Genomic DNA was prepared from the M2 population and the mutation frequency was determined.
The mutation frequency was determined by dividing the total number of mutants by the total number of transformed cells.
We have assumed that some intergenic sequences have derived from coding sequences and could freely accumulate mutations with frequencies determined by the replication-associated mutational pressure.
Considering that MAPK3/1 is downstream of the RAF pathway, we used BRAF mutation frequency to determine a cutoff for p-MAPK3/1 positivity.
Mutation frequencies were determined as previously described [56].
The mutation frequencies were determined for three biological replicates.
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