counts Protocols

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to avoid pitfalls and keep out of trouble in the Neurospora lab. Includes: Culture, incubation, storage, and disposal; Stock preservation; Spore and colony counts; Making crosses and recovering progeny; Use of heterokaryons for allelism tests; Use of unordered asci to estimate the relation of translocation breakpoints to centromeres. - [Read How to Avoid Pitfalls and Keep Out of Trouble in the Neurospora Lab]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The MTT Cell Proliferation Assay measures the cell proliferation rate and conversely, when metabolic events lead to apoptosis or necrosis, the reduction in cell viability. The number of assay steps has been minimized as much as possible to expedite sample processing. The MTT Reagent yields low background absorbance values in the absence of cells. Includes: Determining optimal cell counts, performing an assay, data interpretations and troubleshooting. - [Read MTT Cell Proliferation Assay Protocol]

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles.  Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number).  Total RNA is used as an internal control. - [Read Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer]

Category: Immunology: Immunoassays

Protocol for steroid radioimmunoassay. Includes: SOLVENT DISTILLATION; PREPARATION OF PLASMA SAMPLES; EXTRACTION OF STEROIDS AND COLUMN PACKING; COLUMN CHROMATOGRAPHY; RADIOIMMUNOASSAY; SEPARATION OF BOUND AND FREE COUNTS; DIRECT ASSAYS; SHORT COLUMN CHROMATOGRAPHY. - [Read Steroid Radioimmunoassay Protocol]