Oct 17, 2019
Version 2
Preparation of M9 Media V.2
- 1National University of Singapore
Protocol Citation: NUS iGEM 2019. Preparation of M9 Media. protocols.io https://dx.doi.org/10.17504/protocols.io.8cjhsun
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: October 17, 2019
Last Modified: September 12, 2023
Protocol Integer ID: 28779
Keywords: iGEM
Materials
MATERIALS
Water refers to sterilized deionized water
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
STEP MATERIALS
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
Water refers to sterilized deionized water
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
Protocol materials
Water refers to sterilized deionized water
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
Water refers to sterilized deionized water
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
Water refers to sterilized deionized water
Prepare 5x M9 salt, 1M calcium chloride and magnesium sulfate solutions respectively using the following reagents.
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
Add
200 mL
M9 Minimal Salts 5XMerck MilliporeSigma (Sigma-Aldrich)Catalog #M9956
100 mL
2% casamino
100 µL
1 M Calcium Chloride (CaCl2)Fisher ScientificCatalog #BP510
2 mL
1 Liter Magnesium sulfate solution [1M]G-BiosciencesCatalog #786-530
Top up with sterile deionized water to make a litre of M9 media.
Water refers to sterilized deionized water