Mar 30, 2023
WC Medium
This protocol is a draft, published without a DOI.
- 1University of Konstanz
- Richard W Lambrecht: This protocoll is a digital version of the one used for a long time in the laboratory. It was not developed by the author.
Protocol Citation: Richard W Lambrecht 2023. WC Medium. protocols.io https://protocols.io/view/wc-medium-cr5dv826
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: March 30, 2023
Last Modified: March 30, 2023
Protocol Integer ID: 79749
Keywords: WC medium, Algal culture medium, Algal cultures
Disclaimer
This is a protocol used in the Becks Lab, 2023.
Abstract
Protocoll description to prepare WC medium.
Stock Solution preparation
Stock Solution preparation
The stock solutions don't need to be autoclaved after preparation. Make sure to use volumetric flasks to ensure the right concentrations of the salts and transfer the solution to autoclavable flasks and store it in the fridge. Remember to start the preparation with less than 1L of MilliQ water, and after adding the salts complete the volume to 1L.
Stock solution A: 36,80g of (CaCl2+2H2O) in 1L of MilliQ water
Stock solution B: 37g of (MgSO4+7H2O) in 1L of MilliQ water
Stock solution C: 12,60g of (NaHCO3) in 1L of MilliQ water
Stock solution D: 11,40g of (K2HPO4 +H2O) in 1L of MilliQ water
Stock solution E: 85 g of (NaNO3) in 1L of MilliQ water
Stock solution F: 21,20 g of (Na2SiO3 + 5H2O) in 1L of MilliQ water
Stock solution G (Supplement elements): add the following salts in 1L of MilliQ water:
- 4,36 g of (Na2EDTA)
- 3,15 g of (FeCl3 + 6H2O)
- 0,010 g of (CuSO4 + 5H2O)
- 0,022 g of (ZnSO4 + 7H2O)
- 0,010 g of (CoCl2 + 6H2O)
- 0,180 g of (MnCl2 + 4H2O)
- 0,006 g of (Na2MoO4 + 2H2O)
- 1,0 g of (H3BO3)
Stock solution H (Vitamin Mix): add the following in 1L of MilliQ water:
- Thiamine hydrochloride (Vitamin B1): 0,1 g of (C12H17ClN4OS)
- Biotin: 0,0005 g of (C10H16N2O3S)
- Cyanocobalamin (Vitamin B12): 0,0005 g of (C63H88CoN14O14P)
Aliquot the solution in 15 mL falcon tubes, and freeze them until used at -20 ºC.
Medium preparation (for 1L of WC)
Medium preparation (for 1L of WC)
Weigh 0,115 g of TES Buffer (C6H15NO6S) and add it to a volumetric flask with almost 1L of MilliQ water
Add 1 mL of each of the stock solutions A to G to the medium preparation bottle.
Complete the volume until 1 liter with MilliQ water
Transfer the prepared medium to an autoclavable flask, labeled with autoclaving tape.
On the label put the information about the liquid contained (WC medium), the date, the person responsible for the preparation, and a box to check when adding the solution H (Vitamin Mix).
Autoclave the medium at 121ºC and let it cool down to room temperature.
In a clean bench: syringe filter 1 mL of the defrosted solution H (Vitamin Mix), add it to the medium (1 ml per Liter), and check the box on the tape.