Feb 11, 2023
S. O. C. medium
This protocol is a draft, published without a DOI.
- 1University of Oslo
Protocol Citation: Andreas Sagen 2023. S. O. C. medium. protocols.io https://dx.doi.org/
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 09, 2023
Last Modified: February 11, 2023
Protocol Integer ID: 76690
Keywords: SOB, SOC, Transformation, medium, recovery medium, final step of bacterial cell transformation, bacterial cell transformation, maximal transformation efficiency, transformation, cell
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
GlucoseMerck MilliporeSigma (Sigma-Aldrich)Catalog #G7021
Sodium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9625
AgarMerck MilliporeSigma (Sigma-Aldrich)Catalog #A1296
TryptoneMerck Millipore (EMD Millipore)Catalog #T9410
Yeast ExtractMerck MilliporeSigma (Sigma-Aldrich)Catalog #Y0875
Sodium hydroxide solutionMerck MilliporeSigma (Sigma-Aldrich)Catalog #S8263
Potassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Magnesium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #M0250
500 mL 1 M Glucose solution
Fill the bottle with 400 mL distilled water
Measure 90.08 g Glucose
Materials:
GlucoseMerck MilliporeSigma (Sigma-Aldrich)Catalog #G7021
Add distilled water to a total of 500 mL
Filter sterilize with a 0.2 µm filter
250 mL 250 mM Potassium chloride
Fill the bottle with 200 mL distilled water
Measure 4.66 g Potassium chloride
Materials:
Potassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Add distilled water to a total of 250 mL
Autoclave solution at 121 °C for 00:15:00
15m
125 mL 2 M Magnesium chloride
Fill the bottle with 100 mL distilled water
Measure 50.825 g Potassium chloride
Materials:
Magnesium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #M0250
Add distilled water to a total of 125 mL
Autoclave solution at 121 °C for 00:15:00
15m
1 000 mL S. O. B. medium
Fill the bottle with 950 mL distilled water
Measure 20 g Tryptone, 5 g Yeast extract and 0.5 g Sodium chloride. Add 15 g Agar to create agar plates
Materials:
TryptoneMerck Millipore (EMD Millipore)Catalog #T9410
Yeast ExtractMerck MilliporeSigma (Sigma-Aldrich)Catalog #Y0875
Sodium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9625
AgarMerck MilliporeSigma (Sigma-Aldrich)Catalog #A1296
Add 1 mL Potassium chloride solution (250 millimolar (mM) ) and 0.5 mL Magnesium chloride solution (2 Mass Percent )
Adjust pH to 7.0 with concentrated sodium hydroxide
Materials:
Sodium hydroxide solutionMerck MilliporeSigma (Sigma-Aldrich)Catalog #S8263
Add distilled water to a total of 1000 mL
Autoclave solution at 121 °C for 00:00:00
100 mL S. O. C. medium
Sterilely transfer 98 mL S. O. B. medium to a sterile 100 mL container
Sterilely add 2 mL 1 Mass Percent Glucose solution