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Category Archives: Biology

Biology

PCR Based Moleclar Cloning

In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that it can be easily cloned into a plasmid of interest. You can use similar processes to add overhangs to your…

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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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