{"id":173,"date":"2017-04-19T15:22:15","date_gmt":"2017-04-19T19:22:15","guid":{"rendered":"https:\/\/health.uconn.edu\/mouse-genome-modification\/?page_id=173"},"modified":"2017-04-19T15:22:15","modified_gmt":"2017-04-19T19:22:15","slug":"embryonic-stem-cell-dna-extraction-from-96-well-plates","status":"publish","type":"page","link":"https:\/\/health.uconn.edu\/mouse-genome-modification\/protocols\/embryonic-stem-cell-dna-extraction-from-96-well-plates\/","title":{"rendered":"Embryonic Stem Cell DNA Extraction from 96 Well Plates"},"content":{"rendered":"
    \n
  • Remove media with multiple chanel pipette or by aspiration.<\/li>\n
  • Add 100 \u00b5l PBS, remove.<\/li>\n
  • Add 50 \u00b5l lysis buffer.<\/li>\n
  • Wrap the plate with wet paper towel then seal it in a plastic bag.<\/li>\n
  • Incubate at 50\u00b0C overnight.<\/li>\n
  • Add 100 \u00b5l cold 100 percent etOH.<\/li>\n
  • Leave at room temp (not shaking).<\/li>\n
  • If a plate spinner is available, spin plates for 5\u2019 at 1000 rpm at 10\u00b0C. Pour off EtOH by blotting on paper towel pad.<\/li>\n
  • Wash three times with 150 \u00b5l 70 percent EtOH and pour off.<\/li>\n
  • Pat on paper towel pad until very little liquid comes out.<\/li>\n<\/ul>\n

    Lysis Buffer<\/h2>\n\n\n\n\n\n\n\n\n
    \n

    Final Concentration<\/strong><\/p>\n<\/td>\n

    		\n

    Stock Solution<\/strong><\/p>\n<\/td>\n

    		\n

    10 ml Lysis Buffer<\/strong><\/p>\n<\/td>\n<\/tr>\n

    0.5% Sarcosyl (Sigma L9150)<\/td>\n10%<\/td>\n500 \u00b5l<\/td>\n<\/tr>\n
    200 mM NaCl<\/td>\n5 M<\/td>\n400 \u00b5l<\/td>\n<\/tr>\n
    10 mM EDTA (pH 8.0)<\/td>\n0.5 M<\/td>\n200 \u00b5l<\/td>\n<\/tr>\n
    10 mM Tris.HCl (pH 8.0)<\/td>\n1 M<\/td>\n100 \u00b5l<\/td>\n<\/tr>\n
    1 mg\/ml proteinase K<\/td>\n20 mg<\/td>\n500 \u00b5l<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div><\/div><\/div><\/div><\/div>","protected":false},"excerpt":{"rendered":"

    Remove media with multiple chanel pipette or by aspiration.Add 100 \u00b5l PBS, remove.Add 50 \u00b5l lysis buffer.Wrap the plate with wet paper towel then seal it in a plastic bag.Incubate at 50\u00b0C overnight.Add 100 \u00b5l cold 100 percent etOH.Leave at room temp (not shaking).If a plate spinner is available, spin plates for 5\u2019 at 1000 […]<\/p>\n","protected":false},"author":38,"featured_media":0,"parent":45,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"footnotes":""},"acf":[],"publishpress_future_action":{"enabled":false,"date":"2026-05-06 10:50:19","action":"change-status","newStatus":"draft","terms":[],"taxonomy":""},"_links":{"self":[{"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/173"}],"collection":[{"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/users\/38"}],"replies":[{"embeddable":true,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/comments?post=173"}],"version-history":[{"count":1,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/173\/revisions"}],"predecessor-version":[{"id":174,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/173\/revisions\/174"}],"up":[{"embeddable":true,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/45"}],"wp:attachment":[{"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/media?parent=173"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}