Feb 10, 2023

Public workspaceS. O. C. medium

This protocol is a draft, published without a DOI.
  • 1University of Oslo
Andreas Sagen
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Protocol Citation: Andreas Sagen 2023. S. O. C. medium. protocols.io https://protocols.io/view/s-o-c-medium-cn5svg6e
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 08, 2023
Last Modified: February 10, 2023
Protocol Integer ID: 76690
Keywords: SOB, SOC, Transformation, medium, recovery medium
Abstract
S.O.C. Medium is used in the final step of bacterial cell transformation to obtain maximal transformation efficiency ofE. coli. S.O.C.
Materials
LAF bench
Autoclave
Filter unit
Scale
Protocol materials
ReagentSodium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9625
Step 14
ReagentAgarMerck MilliporeSigma (Sigma-Aldrich)Catalog #A1296
Step 14
ReagentTryptoneMerck Millipore (EMD Millipore)Catalog #T9410
Step 14
ReagentYeast ExtractMerck MilliporeSigma (Sigma-Aldrich)Catalog #Y0875
Step 14
ReagentSodium hydroxide solutionMerck MilliporeSigma (Sigma-Aldrich)Catalog #S8263
Step 16
ReagentPotassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Step 6
ReagentMagnesium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #M0250
Step 10
ReagentGlucoseMerck MilliporeSigma (Sigma-Aldrich)Catalog #G7021
Step 2
500 mL 1 M Glucose solution
500 mL 1 M Glucose solution
Fill the bottle with Amount400 mL distilled water
Measure Amount90.08 g Glucose

Materials:
ReagentGlucoseEmd MilliporeCatalog #G7021
Add distilled water to a total of Amount500 mL
Filter sterilize with a 0.2 µm filter
250 mL 250 mM Potassium chloride
250 mL 250 mM Potassium chloride
Fill the bottle with Amount200 mL distilled water
Measure Amount4.66 g Potassium chloride

Materials:
ReagentPotassium chlorideEmd MilliporeCatalog #P3911
Add distilled water to a total of Amount250 mL
Autoclave solution at Temperature121 °C for Duration00:15:00

15m
125 mL 2 M Magnesium chloride
125 mL 2 M Magnesium chloride
Fill the bottle with Amount100 mL distilled water
Measure Amount50.825 g Potassium chloride

Materials:
ReagentMagnesium chlorideEmd MilliporeCatalog #M0250
Add distilled water to a total of Amount125 mL
Autoclave solution at Temperature121 °C for Duration00:15:00

15m
1 000 mL S. O. B. medium
1 000 mL S. O. B. medium
Fill the bottle with Amount950 mL distilled water
Measure Amount20 g Tryptone, Amount5 g Yeast extract and Amount0.5 g Sodium chloride. Add Amount15 g Agar to create agar plates

Materials:
ReagentTryptoneEmd MilliporeCatalog #T9410
ReagentYeast ExtractEmd MilliporeCatalog #Y0875
ReagentSodium chlorideEmd MilliporeCatalog #S9625
ReagentAgarEmd MilliporeCatalog #A1296
Add Amount1 mL Potassium chloride solution (Concentration250 millimolar (mM) ) and Amount0.5 mL Magnesium chloride solution (Concentration2 Molarity (M) )
Adjust pH to Ph7.0 with concentrated sodium hydroxide

Materials:
ReagentSodium hydroxide solutionEmd MilliporeCatalog #S8263
Add distilled water to a total of Amount1000 mL
Autoclave solution at Temperature121 °C for Duration00:00:00
100 mL S. O. C. medium
100 mL S. O. C. medium
Sterilely transfer Amount98 mL S. O. B. medium to a sterile Amount100 mL container
Sterilely add Amount2 mL Concentration1 Molarity (M) Glucose solution