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FAQ: Why is induced protein insoluble (C2527)?

Check for insolubility - this is important because T7 expression often leads to very high production of protein that can result in the target protein becoming insoluble. Solutions around this are:
  • Induce at a lower temperature (as low as 15°C overnight)
  • Reduce IPTG concentration to 40 µM
  • Induce earlier in growth phase (OD600 = 0.3 or 0.4)
  • Test expression in Lemo21(DE3)