Terrific Broth Base Autoinducible w/o Metals 500 grams/bottle

Terrific Broth Base Autoinducible w/o Metalsis a medium which supports a high cell density and, in the case is formulated for optimum growth of E.coli, maintains growth in the logarithmic phase for a long time. As a result, it yields a greater number of recombinant proteins and plasmic DNA. In some circumstances, it substitutes LB Broth (Lennox) (Cat. 1231) used in genetic studies. Auto Induction Media was firstly formulated and developed by W. studier to grow IPTG-inducible expression strains. The principle of Auto Induction Media is based on carbon sources in the medium that are metabolized differentially to promote high density cell growth and automatically induce protein expression from lac promoters. Auto Induction Media contains glucose as well as lactose as the carbon source. A limited concentration of glucose is metabolized preferentially during growth, which prevents uptake of lactose until the glucose is depleted, usually in mid to late log phase. As the glucose is depleted, lactose can be taken up and converted by the enzyme ß-galactosidase to the inducer allolactose. Allolactose causes the release of lac repressor from its specific binding sites in the DNA and thereby induces expression of T7 RNA polymerase from the lacUV5 promoter and unblocks T7lac promoters, allowing expression of target proteins by T7 RNA polymerase. With Auto Induction Media a high density cell growth is followed by a spontaneous induction of protein expression. There is no need to monitor the cell density and there is no conventional induction with IPTG. Parallel growth of many non-induced or auto-induced cultures is feasible because cultures are simply inoculated and grown to saturation. This is a great convenience and simplifies manual or automated induction and analysis of multiple clones compared to conventional IPTG induction, which requires monitoring growth of each culture and adding inducer at the proper stage of growth. For higher saturation density use the Super Growth Autoinducible Medium w/o Trace Element (Cat. 2095). Tryptone provide nitrogen, vitamins, minerals and amino acids essential for growth. Yeast extract is source of vitamins, particularly the B-group. Potassium phosphates act as a buffer system to prevent cell death. If desired, add a concentration of 0.015 g/l of Trace Elements (Cat. 2109) to supply all of the specific requirements for the bacteria.