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Trump threatens to pull US out of WTO

President Donald Trump has threatened to withdraw the US from the World Trade Organization (WTO), claiming it treats his country unfairly. “If they don’t shape up, I would withdraw from the WTO,” Mr Trump said in an interview with Bloomberg News. The WTO was established to provide rules for global trade and resolve disputes between countries.…

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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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