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A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids
A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lid...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4259245/ https://www.ncbi.nlm.nih.gov/pubmed/25484231 http://dx.doi.org/10.1107/S2053230X14025126 |
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author | Zipper, Lauren E. Aristide, Xavier Bishop, Dylan P. Joshi, Ishita Kharzeev, Julia Patel, Krishna B. Santiago, Brianna M. Joshi, Karan Dorsinvil, Kahille Sweet, Robert M. Soares, Alexei S. |
author_facet | Zipper, Lauren E. Aristide, Xavier Bishop, Dylan P. Joshi, Ishita Kharzeev, Julia Patel, Krishna B. Santiago, Brianna M. Joshi, Karan Dorsinvil, Kahille Sweet, Robert M. Soares, Alexei S. |
author_sort | Zipper, Lauren E. |
collection | PubMed |
description | A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations. |
format | Online Article Text |
id | pubmed-4259245 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-42592452014-12-31 A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids Zipper, Lauren E. Aristide, Xavier Bishop, Dylan P. Joshi, Ishita Kharzeev, Julia Patel, Krishna B. Santiago, Brianna M. Joshi, Karan Dorsinvil, Kahille Sweet, Robert M. Soares, Alexei S. Acta Crystallogr F Struct Biol Commun Laboratory Communications A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations. International Union of Crystallography 2014-11-28 /pmc/articles/PMC4259245/ /pubmed/25484231 http://dx.doi.org/10.1107/S2053230X14025126 Text en © Zipper et al. 2014 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Laboratory Communications Zipper, Lauren E. Aristide, Xavier Bishop, Dylan P. Joshi, Ishita Kharzeev, Julia Patel, Krishna B. Santiago, Brianna M. Joshi, Karan Dorsinvil, Kahille Sweet, Robert M. Soares, Alexei S. A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
title | A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
title_full | A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
title_fullStr | A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
title_full_unstemmed | A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
title_short | A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
title_sort | simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids |
topic | Laboratory Communications |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4259245/ https://www.ncbi.nlm.nih.gov/pubmed/25484231 http://dx.doi.org/10.1107/S2053230X14025126 |
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