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The payoff would come at the end, when they could divide up mutants of interest and launch their own labs to use the tool they'd created.
Chlamydomonas can grow as a haploid and in the absence of a functional photosynthetic apparatus, allowing rapid isolation of mutants of interest.
From the protocols presented, a plasmid-based library of transposon-mutagenized yeast DNA may be used to generate and identify yeast mutants of interest; transposon insertion sites within these mutants may be characterized by vectorette PCR as also described in the chapter.
However, IdealKnock algorithm is a top-down framework that scans for the mutants of interest firstly, followed by the identification of the knockout target by iterative searching process.
Thus, to reduce confounding phenotypes arising from genetic background differences, we first isogenized all Imd pathway mutants of interest.
We first conducted epistatic studies and generated double mutants of interest, including gpb1ssn8, ste20ssn8, cpk1ssn8 and ste12ssn8.
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Traditionally, C. elegans mutations are identified using a combination of multi-point or deficiency mapping by crossing the mutant of interest to marker strains.
A final consideration to keep in mind while using the system is that while the quartet1 mutation can be beneficial, it can also potentially interfere with the results, for example, if the phenotype of the mutant of interest was enhanced or suppressed by qrt1.
One particular seed composition mutant of interest was from the mutant family FN0172932.
It is advisable to cross the mutant of interest to multiple different inbreds (see Fig. 2).
For each mutant of interest, total RNA was extracted from three heterozygote embryos.
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