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Lidstrom:Autoinduction Media: Difference between revisions
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== Inducing media == | == Inducing media == | ||
All of these recipes except MD-5051 are formulated for auto- induction with 0.2 % lactose, which can be replaced for auto- induction with 0.1 % galactose, 0.05 % arabinose, or other sugars subject to glucose inhibition, as appropriate. Lactose and galactose can be exchanged by exchanging 50× 5052 (which provides 0.2 % lactose) and 50× 5051 (which provides 0.1 % galactose) in the recipes for auto-inducing media. Auto-induction of target genes under the con- trol of the T7 lac promoter in BL21-AI requires both arabinose, to induce production of T7 RNA polymerase, and either lactose or galactose, to unblock the T7 lac promoter in the expression plasmid. | |||
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| align="center" style="background:#f0f0f0;"|'''Inducing?''' | | align="center" style="background:#f0f0f0;"|'''Inducing?''' | ||
| align="center" style="background:#f0f0f0;"|'''Purpose''' | | align="center" style="background:#f0f0f0;"|'''Purpose''' | ||
| align="center" style="background:#f0f0f0;"|'''Recipe''' | | align="center" style="background:#f0f0f0;"|'''Recipe''' | ||
| align="center" style="background:#f0f0f0;"|'''Final Concentration''' | |||
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|ZYM-505 || '''inducing''' ||General-Purpose Complex Medium for Rapid Growth to High Density | |||
|| 9.68 ml ZY, 20 μl 1 M MgSO 4 (2 μl 1,000× metals, optional), 100 μl 100× 505, 200 μl 50× M || 1 % N-Z-amine AS, 0.5 % yeast extract, 25 mM Na2HPO4, 25 mM KH2PO4, 50 mM NH 4 Cl, 5 mM Na 2 SO 4 , 2 mM MgSO 4 (0.2× metals, optional), 0.5 % glycerol, 0.05 % glucose. | |||
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| ZYM-5052 || '''inducing''' ||complex auto-inducing medium || 9.57 ml ZY, 20 μl 1 M MgSO 4 (2 μl 1,000× metals, optional), 200 μl 50× 5052, 200 μl 50× M. | |||
|| 1 % N-Z-amine AS, 0.5 % yeast extract, 25 mM Na 2 HPO 4 , 25 mM KH 2 PO 4 , 50 mM NH 4 Cl, 5 mM Na2SO4 , 2 mM MgSO4 (0.2× metals, optional), 0.5 % glycerol, 0.05 % glucose, 0.2 % α-lactose. | |||
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| MDA-5052 || '''inducing''' || defined auto-inducing medium || 9.26 ml water, 20 μl 1 M MgSO 4 , 2 μl 1,000× metals, 200 μl 50× 5052, 40 μl 25 % aspartate, 200 μl 50× M, 280 μl 18aa | |||
|| 25 mM Na 2 HPO 4 , 25 mM KH 2 PO 4 , 50 mM NH 4 Cl, 5 mM Na 2 SO 4 , 2 mM MgSO 4 , 0.2× metals, 0.5 % glycerol, 0.05 % glucose, 0.2 % α-lactose, 0.1 % aspartate, 200 μg/ml of each of 18 amino acids (no C,Y) | |||
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| MDA-5052 || '''inducing''' || defined auto-inducing medium || 9.26 ml water, 20 μl 1 M MgSO 4 , 2 μl 1,000× metals, 200 μl 50× 5052, 40 μl 25 % aspartate, 200 μl 50× M, 280 μl 18aa | |||
|| 25 mM Na 2 HPO 4 , 25 mM KH 2 PO 4 , 50 mM NH 4 Cl, 5 mM Na 2 SO 4 , 2 mM MgSO 4 , 0.2× metals, 0.5 % glycerol, 0.05 % glucose, 0.2 % α-lactose, 0.1 % aspartate, 200 μg/ml of each of 18 amino acids (no C,Y) | |||
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