Protocol describes mutagenesis of yeast with ethyl methane sulfonate (EMS). It causes approximately 40-70% cell death in most haploid laboratory strains, a level of cell killing that is commonly used in mutant hunts with haploid strains. - [Read Ethyl Methane Sulfonate (EMS) Mutagenesis Protocol]
Microneedles attached to micromanipulators are used in the dissection of tetrads, isolation of zygotes from populations of mating haploid cells, and manipulation of individual cells. - [Read Making a Tetrad Dissection Needle Protocol]
Zygotes can be identified by their unique morphology. They can be easily separated away from nonmated cells using a micromanipulator. This method provides an alternative to the selection of diploid cells on a medium that prevents the growth of haploid parent cells. - [Read Picking Zygotes Protocol]
A. thaliana has a very small haploid genome and this makes obtaining DNA somewhat difficult. The most notable problem is that DNA is usually contaminated with polysaccharide which inhibit restriction enzymes as well as other DNA modifying enzymes. This problem is most easily solved by using young plants which have not accumulated as much polysaccharide as older plants. The best results are obtained with plants that are two to three weeks post germinated. - [Read Plant DNA Extraction Protocol]