UltraPure™ TBE Buffer, 10X

No. Autoclaving at temperatures that are too high or pressures that are too high can hydrolyze the agarose polymer and weaken it.

Add the powder to cold buffer while stirring. Let the agarose rehydrate for 1 to 2 hr and then heat slowly until agarose is completely dissolved. Using a microwave at low power settings is acceptable.

One recommendation: get a supply of fabric softener sheets and wipe the bottle with a sheet before opening it.

The precipitation of concentrated TBE stocks may be due to nucleation of salt crystals by dust particles or other insoluble materials. Therefore, filtering the solution through a 0.2 µm cellulose acetate or cellulose nitrate filter after preparation helps avoid this precipitate. [Mayeda A, Krainer A (1991) BioTechniques 10.2, 1820]

Although there is only a very small chance of creating replication competent virus, steps should still be taken to avoid added risk. The virus is expanded in two rounds of amplification, and all secondary expansions should be performed from an initial expansion stock. Secondary amplifications in HEK293 cells should not be performed from the stock of another secondary expansion, as this could increase the chance of making replication competent virus. See the product manual for more detail and guidelines for screening.

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