Fixation is to preserve cells and tissue constituents in as close a life-like state as possible and to allow them to undergo further preparative procedures without change. Fixation arrests autolysis and bacterial decomposition and stabilises the cellular and tissue constituents so that they withstand the subsequent stages of tissue processing. Great detailed guide with protocols. Anthony S-Y Leong. - [Read Fixation and Fixatives - A great guide]
Nice protocol for the preparation of Bacterial cells to allow uptake of plasmid vectors. Dr Peter Kille. Hot Metals Cardiff. - [Read Competent Cell Preparation [CaCl2]]
Formaldehyde cross-linking and chromatin immunoprecipitation assays of
tissue culture cells, Based on Boyd and Farnham. Michelle Kallesen, Rosen Lab. - [Read ChIP Assay Protocol PDF]
Allows students to easily isolate chromosomal DNA from onion cells using the same basic tools and methods that scientists use in the lab. A good introduction to using pipettes. Kate Dollard - [Read DNA Isolation from Onion Cells]
Cell Staining for Immunofluorescence Microscopy. Includes protocols for fixing the cells, Coverslip Preparation for Adherent Cells, Coverslip Preparation for Non-Adherent Cells, Paraformaldehyde Fixation, and Methanol/Acetone Fixation. Blocking protocols include blocking with primary antibody, and incubation with secondary antibody. - [Read Cell Staining for Immunofluorescence Microscopy]
Handbook of Cell Culture. - Sigma Aldrich.- http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_12.html
ECACC Handbook of Cell Culture. Fundamental Techniques in Cell Culture: A Labratory Handbook. Sigma Aldrich. Excellent Guide to Cell Culture, aseptic technique, storage of cell lines, subculture of cells. - [Read Handbook of Cell Culture. - Sigma Aldrich.]
Frozen Competent E.Coli Cells. Methods and protocol for freezing E.Coli Bacteria Competent Cells in aliquots for later use re-use. (Inoue et al., 1990 Gene 96:23). Koshland. - [Read Frozen Competent E.Coli Cells]
Simple Transfection using Calcium chloride and phosphate.Dr. Frank. Arthritis and Immunology Program, Oklahoma City. Medical Research Foundation. - [Read CALCIUM PHOSPATE TRANSFECTION OF HUMAN CELLS]
Cheek cells obtained by rinsing the mouth commercial mouth wash solution. Mouth wash is then discarded into a sterile conical tube and sent to the lab. Based on Salting out procedure. DNA Laboratory, Medical School, Malta. - [Read DNA Extraction from Cheek Cells]
A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.
DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.
Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.
This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.
A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.
Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.