Dec 11, 2024

Public workspace1xBWT buffer V.1

DOI
dx.doi.org/10.17504/protocols.io.eq2lyw99pvx9/v1
  • 1Max Planck Institute for Evolutionary Anthropology;
  • 2The Max Planck Institute for Evolutionary Anthropology
  • MPI EVA Ancient DNA Core Unit
Ancient DNA Core Unit
Icon indicating open access to content
QR code linking to this content
DOI: dx.doi.org/10.17504/protocols.io.eq2lyw99pvx9/v1
Document Citation: Anna Schmidt, Anya Patova, Matthias Meyer 2024. 1xBWT buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2lyw99pvx9/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: 103971
Keywords: capture
Funders Acknowledgements:
Max Planck Society
Grant ID: -
Abstract
Protocol for the preparation of 1xBWT buffer (Maricic et al. 2010), which is used in various protocols of the Ancient DNA Core Unit of the MPI-EVA, especially those involving streptavidin bead binding and washing.

References
Maricic, T., Whitten, M., Pääbo, S. (2010) Multiplexed DNA sequence capture of mitochondrial genomes using PCR products. PLoS One, 16;5(11):e14004. doi: 10.1371/journal.pone.0014004
Note

This protocol describes the preparation of 500 ml buffer.

Materials
Reagent/consumableSupplierCatalogue number
Reagents
Water Sigma Aldrich/Merck1153332500
5 M NaClSigma Aldrich/MerckS5150-1L
1 M Tris-HCl, pH 8.0AppliChemA4577,1000
0.5 M EDTA, ph 8.0AppliChemA4892,1000
Tween-20Thermo Fisher Scientific11417160
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
Water393.75 mL
5 M NaCl100 mL1 M
1 M Tris-HCl, pH 8.05 mL10 mM
0.5 M EDTA, ph 8.01 mL1 mM
100% Tween-200.25 mL0.05%
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).