{"id":1072,"date":"2021-03-29T18:05:43","date_gmt":"2021-03-29T17:05:43","guid":{"rendered":"http:\/\/www.afmb.univ-mrs.fr\/?post_type=facility&#038;p=1072"},"modified":"2024-04-08T16:59:29","modified_gmt":"2024-04-08T15:59:29","slug":"crystallogenesis-and-crystallography","status":"publish","type":"facility","link":"https:\/\/www.afmb.univ-mrs.fr\/en\/facility\/structural-biology\/crystallogenesis-and-crystallography\/","title":{"rendered":"Crystallization and crystallography"},"content":{"rendered":"\n<div class=\"wp-block-group alignfull pageSubHeaderWrapper topSection has-white-color has-dark-blue-background-color has-text-color has-background\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<ul class=\"wp-block-list\">\n<li><a href=\"#presentation\">Introduction<\/a><\/li>\n<\/ul>\n<\/div><\/div>\n\n\n\n<div class=\"wp-block-columns presentationTeamWrapper is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\" id=\"presentation\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\" style=\"flex-basis:66.66%\">\n<p class=\"has-large-font-size\">This <a href=\"https:\/\/www.afmb.univ-mrs.fr\/en\/facility\/structural-biology\/\">PBSIM<\/a> service offers a state-of-the-art automated infrastructure for the screening of hundreds of crystallization conditions from limited amounts of biological samples, and ensures the follow-up of the results.<\/p>\n\n\n\n<p>This service is equipped with a nano-volume crystallization robot for the screening and optimization of conditions; a liquid-handling robot for the filling of reservoirs with screening kit solutions and the custom design of optimization grids; and two cabinets for storage and visualization of the trays (20\u00b0C and 7\u00b0C). Crystal quality and optimal cryo-conditions to be employed are assessed using a complete X-ray diffraction equipment. Data collection of full datasets is also possible.<\/p>\n\n\n\n<p>You can find this service in this video: <a href=\"https:\/\/www.youtube.com\/watch?v=E-5vKfjZoPA&amp;list=PLhk31IqVPY96UXlRAOGLuz_PyGCFnpGDp&amp;index=6\">https:\/\/www.youtube.com\/watch?v=E-5vKfjZoPA&amp;list=PLhk31IqVPY96UXlRAOGLuz_PyGCFnpGDp&amp;index=<\/a><\/p>\n\n\n\n<p>New users of the crystallogenesis service can learn how to use the robots via this video: <a href=\"http:\/\/tutocristallisation.afmb.univ-mrs.fr\/\">Les robots de cristallisation Vd\u00e9f (univ-mrs.fr)<\/a><\/p>\n\n\n\n<div class=\"wp-block-columns is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<p><strong>Notice&nbsp;:<\/strong> The most crucial factor is the quality and purity of your protein sample. We strongly recommend that you check the quality of your protein sample before you plan for an experiment.<\/p>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"800\" height=\"640\" src=\"http:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/image-PF-xtalo-51268.jpg\" alt=\"\" class=\"wp-image-595\" srcset=\"https:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/image-PF-xtalo-51268.jpg 800w, https:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/image-PF-xtalo-51268-300x240.jpg 300w, https:\/\/www.afmb.univ-mrs.fr\/wp-content\/uploads\/2021\/04\/image-PF-xtalo-51268-768x614.jpg 768w\" sizes=\"auto, (max-width: 800px) 100vw, 800px\" \/><\/figure>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-columns tableau is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"network\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"accessibility\">Accessibility<\/h6>\n\n\n\n<p>Academia and industries<\/p>\n<\/div><\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"funding\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"cost\">Cost<\/h6>\n\n\n\n<p>To be defined based on the service requested.<\/p>\n<\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-columns tableau is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"network\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h6 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"how-to-apply\">How to apply?<\/h6>\n\n\n\n<ul class=\"wp-block-list\">\n<li><a rel=\"noreferrer noopener\" href=\"http:\/\/frisbi.eu\/submit-a-proposal\/\" target=\"_blank\">Complete the proposal from<\/a><br>and check \u201cMarseille\u201d for the \u201cHigh throughput crystallization\u201d item<\/li>\n\n\n\n<li><strong>Crystallogenesis contact:<\/strong> <a href=\"https:\/\/www.afmb.univ-mrs.fr\/en\/member\/veronique-zamboni\/\" data-type=\"member\" data-id=\"972\">V\u00e9ronique Zamboni<\/a><\/li>\n\n\n\n<li><strong>Crystallography contact:<\/strong> <a href=\"https:\/\/www.afmb.univ-mrs.fr\/en\/member\/gerlind-sulzenbacher\/\" data-type=\"member\" data-id=\"968\">Gerlind Sulzenbacher<\/a><\/li>\n<\/ul>\n<\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-columns tableau is-layout-flex wp-container-core-columns-is-layout-9d6595d7 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<div id=\"network\" class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h5 class=\"wp-block-heading is-style-default has-orange-color has-text-color\" id=\"specific-equipment\">Specific equipment<\/h5>\n\n\n\n<h6 class=\"wp-block-heading\">Crystallisation<\/h6>\n\n\n\n<ul class=\"wp-block-list\">\n<li>1 Mosquito robot (TTP Labtech)<\/li>\n\n\n\n<li>1 EVO150 (Tecan)<\/li>\n\n\n\n<li>2 RockImager (Formulatrix) cabinets at 20\u00b0C and 8\u00b0C, equipped with UV modules and the CRIMS software<\/li>\n\n\n\n<li>1 incubator at 12-15\u00b0C with no vibrations<\/li>\n<\/ul>\n\n\n\n<h6 class=\"wp-block-heading\">X-ray equipment<\/h6>\n\n\n\n<ul class=\"wp-block-list\">\n<li>1 sealed tube microsource (Primux100, Anton Paar)<\/li>\n\n\n\n<li>1 Mar345 detector mounted on a mardtb stage<\/li>\n\n\n\n<li>1 cryostat (Oxford cryosystems)<\/li>\n<\/ul>\n<\/div><\/div>\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-column columnStickyWrapper is-layout-flow wp-block-column-is-layout-flow\" style=\"flex-basis:33.33%\">\n<div class=\"wp-block-group stickyColumnGroup\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\"><div class=\"singleTeamLeaders clearfix\"><div class=\"singleTeamContactWrapper\"><h5>Team contacts<\/h5><ul><li><a href=\"https:\/\/frisbi.eu\/centers\/afmb\/\" target=\"_blank\"><i class=\"fas fa-link\"><\/i><\/a><\/li><li><a href=\"mailto:gerlind.sulzenbacher@univ-amu.fr\"><i class=\"far fa-envelope\"><\/i><\/a><\/li><li class=\"phoneIconWrapper\"><a class=\"phoneIconLink\"><i class=\"fas fa-tty\"><\/i><\/a><div class=\"phoneNumber\"><a href=\"tel:+33 (0)4 13 94 95 27\">+33 (0)4 13 94 95 27<\/a><\/div><\/li><\/ul><\/div><\/div>\n<\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"wp-block-group accordeon\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h4 class=\"wp-block-heading accordeonTitle\" id=\"publications\">Publications<\/h4>\n\n\n\n<div class=\"wp-block-group accordeonContent\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<ul class=\"wp-block-list\">\n<li>Jabafi I, Selisko B, Coutard B, De Palma AM, Neyts J, Egloff MP, Grisel S, Dalle K, Campanacci V, Spinelli S, Cambillau C, Canard B, Gruez A (2007) Improved crystallization of the coxsackievirus B3 RNA-dependent RNA polymerase. <strong>Acta Crystallogr.<\/strong> F63, 495-8.<\/li>\n\n\n\n<li>Vincentelli R, Bignon C, Gruez A, Canaan S, Sulzenbacher G, Tegoni M, Campanacci V, Cambillau C (2003) Medium-scale structural genomics&nbsp;: strategies for protein expression and crystallization. <strong>Acc. Chem. Res.<\/strong> 36, 165-72.<\/li>\n\n\n\n<li>Sulzenbacher G, Gruez A, Roig-Zamboni V, Spinelli S, Valencia C, Pagot F, Vincentelli R, Bignon C, Salomoni A, Grisel S, Maurin D, Huyghe C, Johansson K, Grassick A, Roussel A, Bourne Y, Perrier S, Miallau L, Cantau P, Blanc E, Genevois M, Grossi A, Zenatti A, Campanacci V, Cambillau C (2002) A medium-throughput crystallization approach. <strong>Acta Crystallogr.<\/strong> D58, 2109-15.<\/li>\n<\/ul>\n<\/div><\/div>\n<\/div><\/div>\n\n\n\n<div class=\"wp-block-group accordeon\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<h4 class=\"wp-block-heading accordeonTitle\" id=\"for-more-information\">For more information<\/h4>\n\n\n\n<div class=\"wp-block-group accordeonContent\"><div class=\"wp-block-group__inner-container is-layout-flow wp-block-group-is-layout-flow\">\n<p>For the preliminary screening step, we use a large range of commercial crystallization kits, including some specific kits, in 96-well plates. For each condition, 100-300 nl of protein solution are mixed with 100 nl of a 100-150 \u00b5l reservoir solution. Usually, we test 3 different protein concentrations (&gt;3 mg\/ml) in varying the volume (100, 200 and 300 nl) of the protein sample. For an initial screening we recommend to assay 2 to 3 different kits. This procedure requires at least&nbsp;70 \u00b5l of protein sample per plate. Various plate types can be provided to users.<\/p>\n\n\n\n<p>For optimization of the initial hits, we have designed a 8&#215;8 grid screen which permits to vary two parameters (e.g., pH value versus precipitant concentration). Each grid requires 10 \u00b5l to 48 \u00b5l of protein sample for the screening of one or three different protein concentrations. In most cases, several rounds of optimization are required before identification of the best conditions. This step can also be performed manually in 24-well Linbro plates.<\/p>\n<\/div><\/div>\n<\/div><\/div>\n","protected":false},"excerpt":{"rendered":"<p>This service provides access to an automated infrastructure for the screening of hundreds of crystallization conditions using very small amounts of biological samples, along with regular observation of the crystallization trays.<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":1060,"menu_order":5,"template":"","class_list":["post-1072","facility","type-facility","status-publish","hentry","entry"],"_links":{"self":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1072","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility"}],"about":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/types\/facility"}],"author":[{"embeddable":true,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/users\/1"}],"version-history":[{"count":20,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1072\/revisions"}],"predecessor-version":[{"id":9252,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1072\/revisions\/9252"}],"up":[{"embeddable":true,"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/facility\/1060"}],"wp:attachment":[{"href":"https:\/\/www.afmb.univ-mrs.fr\/en\/wp-json\/wp\/v2\/media?parent=1072"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}