Oct 21, 2021
Glycerol Stock Preparation for S. elongatus
- 1IISER Pune
- iGEM IISER Pune India 2021
Protocol Citation: Akashdutta 2021. Glycerol Stock Preparation for S. elongatus. protocols.io https://dx.doi.org/10.17504/protocols.io.bzb4p2qw
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 21, 2021
Last Modified: October 21, 2021
Protocol Integer ID: 54364
Abstract
Bacterial cultures in the form of agar plates or liquid cultures are not suitable for long term storage at low temperatures such as 4 degrees. Long-term storage of bacteria is best at temperatures around -80 degrees. However, agar plates and liquid cultures cannot be stored at such temperatures, as the water crystallizes in and around the bacterial cells, rupturing and killing them. Glycerol or DMSO (anti-freezing agents) is added to the media to store samples at such temperatures.
To make n stocks, take an aliquot of the S. elongatus culture such that the OD times the volume is at least 5n.
Centrifuge this aliquot in a falcon at 5000 rpm, 25°C, 00:07:00
7m
Discard the supernatant.
Resuspend the pellet in the minimum amount of BG11 medium required to do so.
Add n mL of 25% glycerol and mix well.
Separate equally into n cryovial tubes.
Store in the stocks in -80 °C .