Dec 11, 2024

Public workspaceHWT buffer V.1

DOI
https://dx.doi.org/10.17504/protocols.io.x54v92kkml3e/v1
  • Anna Schmidt1,
  • Anya Patova1,
  • Matthias Meyer1
  • 1Max Planck Institute for Evolutionary Anthropology
  • MPI EVA Ancient DNA Core Unit
Ancient DNA Core Unit
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DOI: https://dx.doi.org/10.17504/protocols.io.x54v92kkml3e/v1
Document Citation: Anna Schmidt, Anya Patova, Matthias Meyer 2024. HWT buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v92kkml3e/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: 103970
Keywords: multiplexed dna sequence capture of mitochondrial genome, hybridization capture by the ancient dna core unit, multiplexed dna sequence capture, dna analysis of an early modern human, mitochondrial genome, ancient dna core unit of the mpi, ancient dna, dna analysis, hybridization capture, ancient dna core unit, hwt buffer, using pcr product, dna, genome
Funders Acknowledgements:
Max Planck Society
Grant ID: -
Abstract
Protocol for the preparation of HWT buffer (Maricic et al. 2010, 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

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
Troubleshooting
Note

This protocol describes the preparation of 500 ml buffer.

Materials
Reagent/consumableSupplierCatalogue number
Reagents
Water Sigma Aldrich/Merck1153332500
10x AmpliTaq Gold buffer without MgLife Technologies4379874
Tween-20Thermo Fisher Scientific11417160
Consumables
Square media bottle 500 mlVWR391-0630
50 ml serological pipetteCorning BV357550

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
Water449.9 mL
10x AmpliTaq Gold buffer (w/o Mg)50 mL1x
Tween-200.1 mL0.02%


Note
[Labeling]

Label the bottle with the buffer name, batch ID, date and the initials of the person who prepared the buffer. Include lot number of AmpliTaq Gold 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).