Protocol for Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase. - [Read Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes]
Karyotyping is a valuable research tool used to determine the chromosome complement within cultured cells. It is important to keep in mind that karyotypes evolve with continued culture. Because of this evolution, it is important for the interpretation of biochemical or other data, that the karyotype of a specific sub-line be determined. - [Read Karyotyping Protocol]
Media and Culture of Yeast Protocol- https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E5192D47CE9B0776DB7D136C34ED148A&objectid=6673ABA6C2DB2E4E7564EDFA26589256
The yeast, Saccharyomyces cerevisiae, has become an important organism in molecular, biochemical, and genetic analysis. The organism has specific requirements for growth under a variety of conditions. The media, both liquid and solid, simple, define, and complex are describe in this unit. Also included are methods for handling, storing, and shipping stock of yeast. - [Read Media and Culture of Yeast Protocol]
The standard protocol for in situ hybridizations in plants still involves fixing fresh tissue, embedding the tissue in wax, sectioning with a microtome and detection of the transcripts of interest using labeled RNA-probes. This protocol concentrates only on nonradioactive methods, as they are easy to perform, very sensitive and even faster than techniques involving radioisotope labels. - [Read Molecular and Biochemical Analysis of Arabidopsis Protocol]
In this protocol, a bacterial lysogen is constructed from a recombinant bacteriophage {lambda} encoding a fusion protein of interest. The resulting lysogenic colonies are induced to synthesize the fusion protein, which is then isolated in preparation for functional and biochemical analyses. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda} Lysogens]
This protocol describes dissection of yeast tetrads. Tetrad analysis is useful for linkage studies and for constructing strains for genetic and biochemical experiments. - [Read Tetrad Dissection Protocol]
Two distinct modes of cell death, apoptosis and necrosis, can be distinguished on the basis of differences in
morphological, biochemical, and molecular changes occurring in the dying cells. Report the development of flow cytometric techniques that permit concomitant detection of apoptosis and cellular DNA content or BrdU content analysis. - [Read The Use of Flow Cytometry for Concomitant Detection of Apoptosis and Cell Cycle Analysis]
The study of transient gene expression provides a useful complement to the study of stably transformed plants. Transient assays offer a quick method of testing the effects of genes, using either phenotypic, molecular, or biochemical readouts. Transient assays based on Agrobacterium-mediated transformation of leaf explants have been described for other plant species, but it is not known how well these assays work in Arabidopsis. - [Read Transient Expression in Protoplasts]
This protocol describes a method for visualizing early embryo implantation sites using Chicago Sky Blue 6B dye. Once implantation and interimplantation sites are identified and separated, they can be used for cellular, biochemical, and molecular biology analyses. - [Read Visualizing Early Embryo Implantation Sites by Dye Injection Protocol]