Jan 29, 2024
EBT buffer
- 1Max Planck Institute for Evolutionary Anthropology
Document Citation: Anna Schmidt, Sarah Nagel, Matthias Meyer 2024. EBT buffer . protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx3x7zg8j/v1
License: This is an open access document 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
Created: January 15, 2024
Last Modified: April 10, 2024
Document Integer ID: 93532
Funders Acknowledgement:
Max Planck Society
Abstract
EBT buffer (10 mM Tris-HCl, 0.05% Tween-20, pH 8.0) is used in various steps of sample preparation by the Ancient DNA Core Unit of the MPI-EVA.
Note
This protocol describes the preparation of 500 ml buffer.
Materials
| Reagent/consumable | Supplier | Catalogue number | |
| Reagents | |||
| Water | Sigma Aldrich/Merck | 1153332500 | |
| 1 M Tris-HCl, pH 8.0 | AppliChem | A4577, 1000 | |
| Tween-20 | Thermo Fisher Scientific | 11417160 | |
| Consumables | |||
| Square media bottle 500 ml | VWR | 391-0630 | |
| 50 ml serological pipette | Corning BV | 357550 | |
| 5 ml serological pipette | Corning BV | 357543 | |
Equipment
- Automated pipetting aid for glas pipette
Protocol
1. Prepare the buffer in a 500 ml square media bottle by adding the following reagents. Use the glass pipette for transfer of large volumes (> 1 ml). Mix reagents by shaking the bottle.
| Reagent | Volume | Final concentration in reaction | |
| Water | 494.75 ml | ||
| 1 M Tris-HCl, pH 8.0 | 5 ml | 10 mM | |
| Tween-20 | 250 μl | 0.05% | |
| sum | 500 ml |
Note
[Note]
It is also acceptable to use the scale of the bottle to fill up to ~400 ml with water, then adding the remaining 95 ml using the glass pipette.
2. Review the protocol in which the buffer is used to determine whether the buffer should be decontaminated using UV treatment. Instructions for using UV decontamination are provided in the Appendix.
Note
[Labeling]
Label the bottle with the buffer name, batch ID, date and the initials of the person who prepared the buffer.
Attention: Every single bottle prepared at the same day gets a new batch ID. Name the batches with Roman numerals (e.g. batch I, batch II, etc.)
3. Store the buffer at room temperature until used. Shelf life is at least one year from preparation.
Note
[Documentation]
Note the lot numbers, date and initials written on the reagents used for buffer preparation in Labfolder (orange fields).
Appendix
Document
NAME
UV decontamination of materials CREATED BY
Elena Essel
Document
NAME
UV decontamination of reagents/buffersCREATED BY
Elena Essel