Genomic DNA was isolated from overnight cultures of the mutants using the DNeasy Tissue Kit (Qiagen Inc., Valencia, CA).
Essentially, we screened the mutants using the pexophagy assay, as described above, in 96-well plates with fluorescence microscopy.
Chromosomal DNA was obtained from the selected mutants using the GENOME DNA KIT (MP Biomedicals).
Summary: The new version of the TRITON program provides user-friendly graphical tools for modeling protein mutants using the external program MODELLER and for docking ligands into the mutants using the external program AutoDock.
A second list was generated using the same mutants but using the pSDC values instead.
The mutants are used to evaluate the ability of the tests to reveal those artificial faults.
We also managed to rapidly identify the mutation spectrum occurring in the mutant genomes using the output data.
The mutants used for the EMSA (mut-1, -2, and -3) were similar to those employed in the luciferase assay.
The underlined nucleotides were introduced into the mutant constructs, using the QuickChange Mutagenesis kit.
We also analysed the effect of LHS treatment on dmr1 mutants using the spray treatment method.
