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Subsequently, we used these loss-of-function mutants for phenotypic analysis.
Each plasmid was purified by cloning in E. coli and verified by sequencing before being introduced into various yeast mutants for phenotypic characterization (see Materials and methods).
To gain insight into the respective functions of dPGM and iPGM in E.coli, we generated null mutants for phenotypic studies to examine the role of each enzyme.
Bioinformatics algorithms such as SIFT [ 39] and PSSM can be used to prioritize mutants for phenotypic evaluation, as reported before for WKS1 [ 32].
A minimum of 28 embryos per time point is recommended to provide 95% confidence for a 'lethal' call if no live homozygous embryos are identified at a particular stage (as above) and, assuming full penetrance of the phenotype, this number will also be sufficient to achieve at least seven homozygous mutants for phenotypic analysis.
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Amino-acid composition of the C-terminal domain of the Y1F mutant (as described in the 'Materials and methods Construction of the CTD Y1F mutant') used for phenotypic and western blot analyses.
Candidate sem-3 inx-9 sterile mutants were examined for phenotypic resemblance to inx-14 tm2864) inx-14 tm2864ls produced) and fewther characterized.
The results of individual deletions were compared with experimental data for real mutants, for which detailed phenotypic analyses were available.
While screening for phenotypic mutants from MYB-family T-DNA insertion lines, we identified a shoot apex-defective mutant segregating in SALK_074403 from the SALK T-DNA collection [ 26].
When double mutants between pax1-1 (Col) and shy2-2 (L er) were constructed, double mutants lacking the erecta (er) mutation were selected for phenotypic analysis.
F2 founders were then in-crossed to obtain F3 generation homozygote mutants, which were used for phenotypic and genomic analysis.
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