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The following recipe is standardized to 1 L 1. Prepare stock solutions of glucose, MgCl2 and MgSO4 and filter sterilize them a. Glucose 2 M: 18 g glucose in 90 mL of ddH2O, fill up to 100 mL b. MgCl2 2 M: 19 g MgCl2 in 90 mL of ddH2O, fill up to 100 mL c. MgSO4 2 M: 49.3 g MgSO4 x 7 H2O in 90 mL of ddH2O, fill up to 100 mL d.
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20 g. Yeast extract. 5 g. NaCl. 0.5 g. SOC medium is identical to SOB medium, except that it contains 20 m m glucose. To prepare SOB medium, combine the above ingredients and shake until the solutes have dissolved. Add 10 mL of a 250 m m solution of KCl.
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SOB. For solid medium, please see Media Containing Agar or Agarose. Shake until the solutes have dissolved. Add 10 ml of a 250 mM solution of KCl. (This solution is made by dissolving 1.86 g of KCl in 100 ml of deionized H 2 O.) Adjust the pH of the medium to 7.0 with 5 N NaOH (approx. 0.2 ml). Adjust the volume of the solution to 1 liter with.
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Making SOB into SOC: Per 1L of SOB, add 20mL of filter-sterilized 20% glucose solution. CRITICAL Do this under the flow hood. Make sure SOB medium is cooled to <50 C before adding glucose if it is fresh out of the autoclave. If using 50% glucose solution, add 8mL per 1L SOB. We commonly work with 50mL aliquots of SOB and have a 50% stock.
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SOB (Super Optimal Broth) medium is a nutrient-rich microbial growth medium, designed by Douglas Hanahan in 1983. SOB or its glucose-supplemented version SOC is often used during transformation of E. coli competent cells with plasmids (added in transformed cells after heat shock). It results in better survival and higher transformation efficiency as compared to regular LB medium. It contains 2.
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Add SOC to freshly transformed E. coli cells and incubate for up to 1 hour before plating on selective media. 1X SOC Outgrowth Medium: 2% Vegetable Peptone 0.5% Yeast Extract 10 mM NaCl 2.5 mM KCl 10 mM MgCl2 10 mM MgSO4 20 mM Glucose. This product is related to the following categories: Competent Cells Buffers & Diluents Products, Buffers Products
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Just before use, add 5 mL of a sterile solution of 2 m MgCl 2. (This solution is made by dissolving 19 g of MgCl 2 in 90 mL of deionized H 2 O. Adjust the volume of the solution to 100 mL with deionized H 2 O and sterilize by autoclaving for 20 min at 15 psi [1.05 kg/cm 2] on liquid cycle.) For solid medium, please see the recipe entitled.
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Recipe. SOC Medium for E. coli. Reagent Final concentration; Tryptone: 2% (w/v) Yeast extract: 0.5% (w/v) NaCl: 10 m m: KCl: 2.5 m m: MgCl 2: 10 m m: Glucose: 20 m m: Prepare a solution containing the first four reagents, sterilize at 121 °C, and then add sterile MgCl 2 and glucose.
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Super Optimal Broth (SOB medium) is a nutrient-rich bacterial growth medium used for microbiological culture, generally of Escherichia coli.This nutrient-rich microbial broth contains peptides, amino acids, water soluble vitamins and glucose in a low-salt formulation. It was developed by Douglas Hanahan in 1983 and is an adjusted version of the commonly used LB medium (lysogeny broth).
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Soc medium preparation: Used in bacterial transformation. Ingredients for SOB: 0.5% (w/v) yeast extract 2% (w/v) tryptone 10 mM NaCl 2.5 mM KCl 20 mM MgSO 4 Per liter: 5 g yeast extract 20 g tryptone 0.584 g NaCl 0.186 g KCl 2.4 g MgSO 4 Note: Some formulations of SOB use 10 mM MgCl 2 and 10 mM MgSO 4
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S.O.C. medium has the same formulation as the rich S.O.B. (Super Optimal Broth) medium with additionally added 10 mM MgCl 2 and 20 mM glucose (as published by Hanahan in 1983). S.O.C. Medium is supplied as 10 x 10 mL bottles of ready-to-use liquid medium. Each lot of S.O.C. Medium is tested to ensure conformance with the most current approved.
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Step 3: Cool down the medium to room temperature. Add 2.5 ml of 1M KCl and mix. Adjust the pH 7.0 with 5N NaOH (~0.2 ml). Note:The SOB medium is not very highly buffered. The pH of the medium drops, as the growing culture reaches the near saturation phase. Step 4: Adjust the volume to 960 ml with deionized / Milli-Q water.
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SOC Medium, shorthand for Super Optimal broth with Catabolite repression, is a nutritionally rich bacterial culture medium. The recipe is differs slightly with LB broth, containing 20 g/L tryptone, 5 g/L yeast extract, and 0.5 g/L sodium chloride. A defining ingredient is 20 mM glucose, which must be added separately after the media has cooled.
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9. ( Super Optimal broth with Catabolite Repression) SOC media contains glucose whereas Luria Broth (LB) doesn't. SOB also has a more delicate salt balance. Naturally the glucose will help the cells recover and grow faster after the heat shock. Your transformation efficiency should improve since less cells will die.
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This medium allows the transformed cells to recover optimally. In the SOC medium, tryptone and yeast extract provide E. coli with a source of carbon and nitrogen, in addition, the medium contains minerals, vitamins and amino acids. Sodium chloride and potassium chloride in the medium maintain ion transport and osmotic balance.
