Feb 06, 2023

Public workspaceInoue transformation buffer

This protocol is a draft, published without a DOI.
Inoue transformation buffer
  • Andreas Sagen1
  • 1University of Oslo
Andreas Sagen
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Protocol Citation: Andreas Sagen 2023. Inoue transformation buffer. protocols.io https://protocols.io/view/inoue-transformation-buffer-cnvbve2n
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 05, 2023
Last Modified: February 06, 2023
Protocol Integer ID: 76419
Keywords: Transformation, bacteria, chemical, Inoue, chemical transformation method of prokaryote, inoue method of transformation, such as the classic calcium chloride transformation method, chemical transformation method, classic calcium chloride transformation method, inoue transformation, transformation, inoue method, prokaryote, buffer, calcium chloride, significant improvements to method, method
Abstract
The Inoue method of transformation is a chemical transformation method of prokaryotes (bacteria), which add significant improvements to methods such as the classic Calcium chloride transformation method.
Materials
LAF
Scale
Filter unit
Protocol materials
ReagentManganese(II) chloride tetrahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #M3634
ReagentCalcium chloride dihydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #C3881
ReagentPIPESMerck MilliporeSigma (Sigma-Aldrich)Catalog #P1851
ReagentPotassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Troubleshooting
10x Inoue transformation buffer
Add Amount80 mL distilled water to a Amount100 mL tube
Measure and add Amount10.89 g Manganese chloride, Amount2.2 g Calcium chloride, Amount3.0 g PIPES and Amount18.65 g Potassium chloride

Materials:
ReagentManganese(II) chloride tetrahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #M3634
ReagentCalcium chloride dihydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #C3881 ReagentPIPESMerck MilliporeSigma (Sigma-Aldrich)Catalog #P1851
ReagentPotassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Add distilled water to Amount100 mL
Filter sterilize with a 0.2 µm pore-size filter and store frozen at Temperature-20 °C