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Category Archives: Biology

Biology

Protocol for Zymogram

Reagents: 1% Gelatin in H2O (Fisher Blood 275) 1% Casein (Sigma) SDS-PAGE gel stock w/o urea Wash buffer: 2.5% Triton X-100 in H2O (+0.02% NaN3) Incubation Buffer: 50mM Tris-HCl (PH8.0), 5mM CaCl2, 0.02% NaN3 Gels: Regular separating gel containing 10-12% substrate Regular stacking gel Protocol: 1- Collect media from cells (if desired, inactivate non-metalloproteases with…

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Making mutants in Dickeya and Pectobacterium species

Objective: To disrupt a gene in a soft rot bacterial genome. Insertion mutation: an antibiotic resistance gene cassette is inserted into a convinient restriction site in the gene. One major pitfalls with this type of mutant is that the remaining N-terminal portion of the protein is sometimes functional and can even have dominant negative effects.…

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Citrate test

Citrate test

Principle: The citrate test detects the ability of an organism to use citrate as the sole source of carbon and energy. Bacteria are inoculated on a medium containing sodium citrate and a pH indicator such as bromothymol blue. The medium also contains inorganic ammonium salts, which are utilized as sole source of nitrogen. Use of citrate…

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B and T Cells Isolation

A serial of protocols for isolation and purification of B cells and T cells. This is a collection of internet based bioprotocols.   B Cell Isolation   Dynal Biotech (Invitrogen) – Dynabeads ( T Cell Isolation Procedure)   Fractionation of T and B Cells Using Magnetic Beads Protocol –   Isolation of Ly-1+/CD5+ B Cells…

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A Guide to Bacteria Preservation 

Bacteria Preservation:  Refrigeration, Freezing and Freeze Drying Between stock cultures, mutant strains, and genetically engineered variants, the number of individual bacterial cultures which any one lab can accumulate can be numerous.  Indeed, the number of variations created in the process of engineering one plasmid can be astounding.  And most labs will hold on to all…

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