{"id":6641,"date":"2015-11-23T22:12:06","date_gmt":"2015-11-24T03:12:06","guid":{"rendered":"http:\/\/admin.acceleratingscience.com\/?p=6641"},"modified":"2016-04-25T18:26:33","modified_gmt":"2016-04-25T18:26:33","slug":"tips-for-transfection","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/cellculture\/tips-for-transfection\/","title":{"rendered":"Tips for Transfection"},"content":{"rendered":"

Transfection is a powerful tool with a range of applications in molecular and cell biology, particularly in studying gene expression for drug discovery. Though there are many different transfection techniques, you can boost the reproducibility and reliability of your results by following these general practices:<\/p>\n

    \n
  1. \n

    \u00a0\u00a0\u00a0\u00a0 Planning is paramount<\/h2>\n<\/li>\n<\/ol>\n

    Before you begin, familiarize yourself with the transfection protocol and make sure you have everything you need, including sufficient reagents, DNA and media.<\/p>\n

      \n
    1. \n

      \u00a0\u00a0\u00a0\u00a0 Start with healthy cells<\/h2>\n<\/li>\n<\/ol>\n

      To give your transfection the highest chance of success, only use cells with over 90% viability. If your cells are frozen, make sure you passage them 3-4 times after thawing to give them time to recover and adjust to their normal rate of growth. If cells aren\u2019t passaged regularly, they can overgrow, which can affect the outcome of transfection significantly.<\/p>\n

        \n
      1. \n

        \u00a0\u00a0\u00a0\u00a0 Use high-quality DNA<\/h2>\n<\/li>\n<\/ol>\n

        In order to achieve reliable results, it\u2019s crucial to use only the purest DNA. Purity can be measured by the OD 260\/280 ratio, and should generally be between 1.7-1.8 \u2013 anything higher or lower could indicate contaminants and shouldn\u2019t be used.<\/p>\n

          \n
        1. \n

          \u00a0\u00a0\u00a0\u00a0 Plating your cells<\/h2>\n<\/li>\n<\/ol>\n

          Cell plating hugely impacts transfection efficiency, so it\u2019s important to get this step right. Plate your cells no sooner than the day before you plan to transfect, aiming for 70-90% confluency at the time of transfection.<\/p>\n

            \n
          1. \n

            \u00a0\u00a0\u00a0\u00a0 Prepare lipid DNA complexes<\/h2>\n<\/li>\n<\/ol>\n

            Lipid DNA complexes have a positively charged surface that aids the delivery of nucleic acids into cells. For optimal fusion, dilute both the lipid and DNA in medium separately, before adding them together to form your complex.<\/p>\n

              \n
            1. \n

              \u00a0\u00a0\u00a0\u00a0 Use controls to assess efficiency<\/h2>\n<\/li>\n<\/ol>\n

              Following the procedure, you can measure the number of transfected cells against a positive control such as GFP or Lac7 reporter plasmid by microscopy or flow cytometry. Expression should be detectable approximately 24 to 48 hours following transfection.<\/p>\n

              The next time you use transfection in your research, we recommend incorporating the above practices into your workflow to make sure your transfected cells have optimal expression.<\/p>\n

              Next Steps<\/h2>\n

              Looking for additional transfection tips? Find more valuable insights here<\/a>.<\/p>\n","protected":false},"excerpt":{"rendered":"

              Transfection is a powerful tool with a range of applications in molecular and cell biology, particularly in studying gene expression for drug discovery. Though there are many different transfection techniques, you can boost the reproducibility and reliability of your results by following these general practices: \u00a0\u00a0\u00a0\u00a0 Planning is paramount Before you begin, familiarize yourself with<\/p>\n","protected":false},"author":130,"featured_media":8440,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[4],"tags":[58,59,60],"division":[],"class_list":{"0":"post-6641","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-general-cellculture","8":"tag-tranfection-reagent","9":"tag-transfection-techniques","10":"tag-transfection-tips","11":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"\nTips for Transfection - Advancing Cell Culture<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.thermofisher.com\/blog\/cellculture\/tips-for-transfection\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Tips for Transfection\" \/>\n<meta property=\"og:description\" content=\"Transfection is a powerful tool with a range of applications in molecular and cell biology, particularly in studying gene expression for drug discovery. 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