Aug 05, 2019
Version 2
Make LB agar medium V.2
- Gurdon Institute mediak1
- 1Wellcome Trust / Cancer Research UK Gurdon Institute
Protocol Citation: Gurdon Institute mediak 2019. Make LB agar medium. protocols.io https://dx.doi.org/10.17504/protocols.io.55zg876
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: August 05, 2019
Last Modified: August 05, 2019
Protocol Integer ID: 26521
Abstract
Make LB agar medium.
Materials
MATERIALS
LB-Broth Miller (= LB mix)FormediumCatalog #LMM0104
LBMixAndAgar5L.xls | Solution | LB (ready mix) + Agar | |||||
| Strength | pH: | 7.0 | Batch size | 5L | ||
| Ingredients | Quantity | |||||
| LB mix | 125g | |||||
| NaOH 10M | few drops | |||||
| Distilled H2O | up to 5L | |||||
| Agar | per 1L bottle | 14g | ||||
| per 500ml bottle | 7g | |||||
| Method | ||||||
| 1 | In the fume cupboard, add the LB mix to 2L distilled H2O in a bell jar | |||||
| 2 | Transfer the bell jar to a magnetic stirrer on the bench | |||||
| 3 | Add distilled H2O to approx. 4L mark using a jug | |||||
| 4 | Allow the solutes to dissolve then adjust pH to 7.0 using 10M NaOH | |||||
| 5 | Adjust final volume to 5L using distilled H2O | |||||
| 6 | Aliquot (using a pump), label & date | |||||
| 7 | Dispense 14g agar and a magnetic flea to each 1L bottle (OR 7g agar and a magnetic flea to each 500ml bottle) | |||||
| 8 | Autoclave | |||||