Feb 06, 2023
Inoue transformation buffer
This protocol is a draft, published without a DOI.
- 1University of Oslo
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
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
Potassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Step 2
Manganese(II) chloride tetrahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #M3634
Step 2
Calcium chloride dihydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #C3881
Step 2
PIPESMerck MilliporeSigma (Sigma-Aldrich)Catalog #P1851
Step 2
10x Inoue transformation buffer
10x Inoue transformation buffer
Add 80 mL distilled water to a 100 mL tube
Measure and add 10.89 g Manganese chloride, 2.2 g Calcium chloride, 3.0 g PIPES and 18.65 g Potassium chloride
Materials:
Manganese(II) chloride tetrahydrateSigma AldrichCatalog #M3634
Calcium chloride dihydrateSigma AldrichCatalog #C3881 PIPESSigma AldrichCatalog #P1851
Potassium chlorideSigma AldrichCatalog #P3911
Add distilled water to 100 mL
Filter sterilize with a 0.2 µm pore-size filter and store frozen at -20 °C