{"id":279,"date":"2023-07-07T12:37:56","date_gmt":"2023-07-07T04:37:56","guid":{"rendered":"http:\/\/yuhongtaolab.com\/?post_type=resource&#038;p=279"},"modified":"2023-07-07T12:37:56","modified_gmt":"2023-07-07T04:37:56","slug":"lipofactemine-2000-transfection-protocol","status":"publish","type":"resource","link":"http:\/\/yuhongtaolab.com\/en\/resource\/lipofactemine-2000-transfection-protocol\/","title":{"rendered":"Lipofactemine 2000 transfection protocol"},"content":{"rendered":"<p><strong>Transfection in 6-well plate<\/strong><br \/>\nSeed cells in 6-well plate, seed ~1e6\/well and culture for overnight.<br \/>\nPrepare transfection mixture as below:Dilute 10ul Lipofectamine 2000 in 125ul Opti-MEM, mix well and incubate at RT for 5min.<br \/>\nDilute 2.5ug plasmid in 125ul Opti-MEM, mix well and incubate at RT for 5min.<br \/>\nAdd diluted Lipofectamine to diluted plasmid, mix well and incubate at RT for 10min.<br \/>\nChange 1.5ml\/well serum free medium (or Opti-MEM)<br \/>\nAdd transfection mixture onto cells.<br \/>\n4-5h after transfection, change 2ml\/well complete medium.<\/p>\n<p><strong>Notes<\/strong><br \/>\nThe amount of transfected plasmid can be varied from 2ug to 4ug.<br \/>\nThe amount of Lipofectamine 2000 reagent can be varied from 5ul to 12ul.<br \/>\nThe amount of Opti-MEM can be veried from 125ul to 250ul. (final transfection mixture volume 250ul to 500ul).<br \/>\nThe transfection mixture must be made in Opti-MEM, but the mixture can be added directly to cells in culture medium with serum and antibiotic.<\/p>","protected":false},"featured_media":0,"template":"","class_list":["post-279","resource","type-resource","status-publish","hentry"],"acf":[],"_links":{"self":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/resource\/279","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/resource"}],"about":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/types\/resource"}],"wp:attachment":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/media?parent=279"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}