Mononuclear phagocyte progenitor cells derived from femoral and tibial bone marrow are propagated in the presence of M-CSF. This macrophage growth factor is secreted by L929 cells and is used in the form of L929 cell conditioned medium. The progenitor cells proliferate and differentiate through monoblast, promonocyte and monocyte stages before maturing to macrophages. At this time the cells become firmly adherent to the culture vessel. - [Read Isolation and Culture of Mouse Bone Marrow-Derived Macrophages Protocol]
Protocol describes a method for transformation of the Tetrahymena using electroporation. The vector is electroporated into cells after mating, where it is incorporated into the DNA of developing macronuclei. Because T. thermophila can be propagated indefinitely without conjugation, transformation of the macronucleus provides a way to obtain stable somatic transformants. DNA vectors transformed using this protocol include those containing drug-resistant versions of Tetrahymena genes. - [Read Transformation of Tetrahymena thermophila by Electroporation Protocol]
Protocol describes a method for transformation of the Tetrahymenausing electroporation. The vector is electroporated into cellsafter mating, where it is incorporated into the DNA of developingmacronuclei. Because T. thermophila can be propagated indefinitelywithout conjugation, transformation of the macronucleus providesa way to obtain stable somatic transformants. - [Read Transformation of Tetrahymena thermophila by Electroporation Protocol]
Protocol describes the general procedure for creating mutations in the DNA of Drosophila by exposure to X-rays. Irradiation of cells with X-rays creates double strand breaks (DSBs) in DNA. Mutations introduced in the DNA of germ line cells (sperm) are propagated by mating the exposed males to virgin females. The progeny of this cross can be mated to each other so that a percentage of the subsequent offspring will have two copies of the same mutant allele. - [Read X-Ray Mutagenesis of Drosophila Protocol]