Dec 11, 2024

Public workspaceWash buffer 1 V.1

DOI
dx.doi.org/10.17504/protocols.io.ewov1955ylr2/v1
  • 1Max Planck Institute for Evolutionary Anthropology
  • MPI EVA Ancient DNA Core Unit
Ancient DNA Core Unit
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DOI: dx.doi.org/10.17504/protocols.io.ewov1955ylr2/v1
Document Citation: Anna Schmidt, Anya Patova, Matthias Meyer 2024. Wash buffer 1. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1955ylr2/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: July 24, 2024
Last Modified: December 11, 2024
Document Integer ID: 103969
Funders Acknowledgements:
Max Planck Society
Grant ID: -
Abstract
Protocol for the preparation of Wash buffer 1 (Fu et al. 2013), which is used in hybridization capture by the Ancient DNA Core Unit of the MPI-EVA.

References
Fu, Q., Meyer, M., Gao, X., Stenzel, U., Burbano, H.A., Kelso, J., Pääbo, S. (2013) DNA analysis of an early modern human from Tianyuan Cave, China. Proc Natl Acad Sci U S A. 110(6):2223-7. doi: 10.1073/pnas.1221359110
Note

This protocol describes the preparation of 500 ml buffer.

Materials
Reagent/consumableSupplierCatalogue number
Reagents
Water Sigma Aldrich/Merck1153332500
20x SSC bufferAmbionAM9770
20 % SDS solutionServa39575.01
Consumables
Square media bottle 500 mlVWR391-0630
50 ml serological pipetteCorning BV357550
5 ml serological pipetteCorning BV357543

Equipment
  • Serological pipette controller (e.g. battery-powered pipetting aid ROTILABO, cat. no. TC16.1)

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.
ReagentVolumeFinal concentration in reaction
Water472.5 mL
20x SSC buffer25 mL1x
20% SDS solution2.5 mL0.1%
sum500 ml


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.)

2. 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).