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文献和实验Streptomyces:Protocols/Mini-Maxi Prep
) except for the phenol/chloroform step(s) and the RNase incubation step (obviously!). Spins should be done at the maximum capacity of the centrifuge/tubes. A mini prep uses 3mL of cells; a maxi prep uses 50mL of cells (different steps are indicated in bold ). Cell
. Sponge - filter paper - gel - membrane - filter paper - sponge 3. Transfer for 1 hr at 1 amp at 4°C on a stir plate. Bigger proteins might take longer to transfer. For the Mini-Transblot, it's 100 V for 1 hr with the cold pack and prechilled buffer
Overview Blotting for large V5-tagged proteins in S. cerevisiae Materials Pierce ) Halt EDTA-free Protease Inhibitor (Pierce ) NuPAGE Novex Bis-Tris 4-12% mini gels (Invitrogen NP0321BOX) Prestained
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