Jun 25, 2020
Making Freezing Solution
- 1Imperial College London
- Behavioural Genomics
Protocol Citation: Priota Islam 2020. Making Freezing Solution. protocols.io https://dx.doi.org/10.17504/protocols.io.bhwgj7bw
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: June 25, 2020
Last Modified: June 25, 2020
Protocol Integer ID: 38568
Abstract
The freezing solution can be made in advance and stored in sterile containers at room temperature till needed.
Steps
Steps
Book the autoclave beforehand
Make 0.05M K2HPO4 by dissolving 8.71g of K2HPO4 in 1L of DIW
Make 0.05M KH2PO4 by dissolving 6.80g of KH2PO4 in 1L of DIW
Make S Buffer solution by adding the following:
129 ml 0.05 M K2HPO4
871 ml 0.05 M KH2PO4
5.85 g NaCl
Make the freezing solution by adding the S buffer to 30% glycerol (v/v):
Just add 700ml of S buffer to 300ml of glycerol to get the desired concentration
Autoclave the mixture. (Aliquoting first is not advised due to handling issues upon adding glycerol)
After autoclaving, aliquot into sterile 200ml bottles labelled with the name and date of the solution prepared.
Notes
Notes
Before using the freezing solution always check the bottle for any growth/contaminants
Discard the bottle if you see any contaminants and open a new one
Aliquot into falcon tubes from 200ml bottles to reduce contamination. Discard the tube after freezing do not put the contents back to the 200ml bottle!