Laboratory Notes

Overview: Resuspension buffer is used to resuspend the harvested bacterial cells from the culture during plasmid isolation by the alkaline…

Overview The neutralization solution (solution III) is used for the isolation of plasmid DNA by the alkaline lysis method. The…

Overview Lysis solution (solution II) is used for the isolation of plasmid DNA by the alkaline lysis method. The plasmid-containing…

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EDTA (EthyleneDiamineTetraAcetic acid), a polyamino carboxylic acid, is extensively used in molecular biology experiments as a chelating agent. It sequesters…

Overview Tris is also known as THAM, Tris base, Trizma, Tris(hydroxymethyl)aminomethane. Tris is one of the most frequently used buffers…

Overview Bovine pancreatic RNase (RNase A) is an endoribonuclease. It specifically degrades single-stranded RNA at C and U residues at…

A 50x TAE buffer can be prepared by mixing and dissolving 242 g Tris base, 100 ml of 0.5 M EDTA and 57.1 ml glacial acetic acid in a deionized water to a final volume of 1000 ml. The pH of the final solution should be between 8.2 – 8.4. 

TBE is a very common electrophoresis buffer for DNA agarose gel electrophoresis. It is prepared as a 5X stock solution that can be diluted to 1x or 0.5x working solution for DNA agarose gel electrophoresis. The 5X concentrated solution contains 0.445 M Tris borate and 0.01 M EDTA (pH 8.2 – 8.4).

A 5X stock solution is prepared by dissolving 54 g Tris base, 27.5 g Boric acid, and 20 ml of 0.5 M EDTA in water to a final volume of 1000 ml. The solution is sterilized by autoclaving and is stored at room temperature.

Overview Tetracyclines are a group of broad-spectrum antibiotics. They are active against a wide range of microorganisms including bacteria (both…

Overview   CAS Number 9001-99-4 EC Number 232-646-6 Enzyme Commission (EC) Number 3.1.27.5 Synonyms Pancreatic Ribonuclease, RNase A, Ribonucleate 3′-pyrimidinooligonucleotidohydrolase,…

Overview Streptomycin function by inhibiting protein synthesis in bacteria. Binding of streptomycin to the small 16S rRNA of the 30S…

Overview Kanamycin is a water-soluble, aminoglycoside bactericidal antibiotic. It was originally purified from the bacterium Streptomyces kanamyceticus. It inhibits protein…

LB medium is a rich growth medium, commonly used to grow E. coli. LB, Lennox Broth contains 1% Bacto-tryptone, 0.5% Bacto-yeast extract and 0.5% Sodium chloride. A 1000 ml LB, Lennox Broth is prepared by dissolving 10 g Bacto-tryptone, 5 g Bacto-yeast extract and 5 g sodium chloride in deionized water to a final volume of 1000 ml. After adjusting the pH 7.0 with 5N NaOH medium is autoclaved and stored in a cold room.

OVERVIEW Luria-Bertani (LB) medium is a rich medium, very commonly used to grow E. coli. It contains tryptone, yeast extract,…