Soc Medium Recipe Besto Blog
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 ยฐC, and then add sterile MgCl 2 and glucose.
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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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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.
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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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Super optimal medium with catabolic repressor (SOC) medium is a glucose-rich microbial growth medium used primarily in the recovery step of Escherichia coli competent cell transformations. The culture of E.coli cells in this nutrient-rich microbial broth increases the transformation efficiency of recombinant plasmids containing toxic protein genes by inhibiting the unintended activation of the.
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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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Table Of Contents Introduction SOC (Super Optimal broth with Catabolite repression) is a variant of SOB media. SOC is a nutrient-rich culture media developed by Douglas Hanahan in 1983. SOC is typically used for bacterial transformation. Since it is nutrient-rich media, it helps bacteria to recover faster after heat shock or electroporation. Applications SOC medium is optimized for growing.
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SOC Broth is a rich media used primarily in the recovery step of Escherichia coli competent cell transformations. Use of SOC Broth maximizes the transformation efficiency of competent cells. Directions per Liter: Dissolve 34.46g in 800-900ml of ddH2O stirring gently with heating until completely solubilized. Adjust pH of the medium to the.
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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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2.5 mM KCl. 10 mM MgCl2. 10 mM MgSO4. 20 mM Glucose*. *Note: add Glucose after autoclaving the solution with the remaining ingredients, and letting it cool down. Sterilize the final solution by passing it through a 0.2 ยตm filter. SOC medium can be stored at room temperature and is stable for several years. icon_0011_idea-s.
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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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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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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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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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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