{"experiment":{"id":436,"name":"Transformation of E. coli with electroporation","description":"This template will show you how to transform E.coli bacteria with electroporation. In molecular biology, transformation can be artificially induced through the creation of pores in the bacterial cell walls. \r\nElectroporation is the use of high-voltage electric shocks to introduce DNA into cells. It can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression and, because it requires fewer steps, can be easier than alternate techniques.","project_id":223,"created_by_id":202,"last_modified_by_id":202,"archived":false,"archived_by_id":null,"archived_on":null,"restored_by_id":null,"restored_on":null,"created_at":"2018-11-13T09:57:55.843Z","updated_at":"2019-02-07T14:41:26.829Z","uuid":"59b17955-fba6-405a-a6b8-6a78a05af66f"},"my_modules":[{"my_module":{"id":2166,"name":"Analysis of transformation","due_date":null,"description":null,"x":67,"y":7,"my_module_group_id":1165,"created_at":"2018-11-13T15:51:41.812Z","updated_at":"2018-12-13T15:38:36.161Z","archived":false,"archived_on":null,"created_by_id":202,"last_modified_by_id":202,"archived_by_id":null,"restored_by_id":null,"restored_on":null,"nr_of_assigned_samples":0,"workflow_order":3,"experiment_id":436,"state":"uncompleted","completed_on":null},"outputs":[],"my_module_tags":[],"task_comments":[],"my_module_repository_rows":[],"user_my_modules":[],"protocols":[{"protocol":{"id":3508,"name":null,"authors":null,"description":null,"added_by_id":null,"my_module_id":2166,"team_id":1,"protocol_type":"unlinked","parent_id":null,"parent_updated_at":null,"archived_by_id":null,"archived_on":null,"restored_by_id":null,"restored_on":null,"created_at":"2018-11-13T15:51:41.820Z","updated_at":"2019-02-20T07:58:02.380Z","published_on":null,"nr_of_linked_children":0},"protocol_protocol_keywords":[],"steps":[{"step":{"id":5189,"name":"Obtaining single colonies","description":"\u003c!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\"\u003e\n\u003chtml\u003e\u003cbody\u003e\n\u003cp\u003eFor each electroporation, \u003cstrong\u003e2.5, 25\u003c/strong\u003e and \u003cstrong\u003e250 pl\u003c/strong\u003e of cells, each in duplicate, were spread on LB plates containing \u003cstrong\u003e12.5 mg/mL\u003c/strong\u003e chloramphenicol. For transformations of ligations with the vector, plates contained \u003cstrong\u003e50 µg/mL\u003c/strong\u003e X-gal and \u003cstrong\u003e25 µg/mL\u003c/strong\u003eIPTG.\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eGrowth conditions:\u003c/strong\u003e\u003c/p\u003e\n\u003cul\u003e\n\u003cli\u003e37°C\u003c/li\u003e\n\u003cli\u003e24 hours\u003c/li\u003e\n\u003c/ul\u003e\n\u003cbr\u003eScoring of colonies.\u003c/body\u003e\u003c/html\u003e","position":0,"completed":false,"completed_on":null,"user_id":202,"created_at":"2018-12-13T15:04:47.190Z","updated_at":"2019-02-19T14:21:06.329Z","last_modified_by_id":202,"protocol_id":3508},"checklists":[],"step_comments":[],"step_assets":[],"assets":[],"step_tables":[],"tables":[]},{"step":{"id":5190,"name":"Analysis","description":"\u003c!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\"\u003e\n\u003chtml\u003e\u003cbody\u003e\n\u003cp\u003eThere are many factors that caninfluencetransformation efficiency. Therefore after each transformation, an assessmentof efficiencyis needed.\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eFor gene expression common methods used:\u003c/strong\u003e\u003c/p\u003e\n\u003col\u003e\n\u003cli\u003eFlow Cytometry\u003c/li\u003e\n\u003cli\u003eqPCR\u003c/li\u003e\n\u003c/ol\u003e\n\u003cstrong\u003eFor protein expression common methods are:\u003cbr\u003e\u003c/strong\u003e\u003cbr\u003e\n\u003col\u003e\n\u003cli\u003eWestern blot analysis (\u003ca href=\"https://www.protocols.io/view/12-sds-page-western-blot-rwrd7d6\" target=\"_blank\" rel=\"noopener\"\u003eClickhere\u003c/a\u003e)\u003c/li\u003e\n\
