{"id":186,"date":"2017-04-19T15:30:55","date_gmt":"2017-04-19T19:30:55","guid":{"rendered":"https:\/\/health.uconn.edu\/mouse-genome-modification\/?page_id=186"},"modified":"2017-04-19T15:30:55","modified_gmt":"2017-04-19T19:30:55","slug":"tail-dna-extraction","status":"publish","type":"page","link":"https:\/\/health.uconn.edu\/mouse-genome-modification\/protocols\/tail-dna-extraction\/","title":{"rendered":"Tail DNA Extraction"},"content":{"rendered":"
    \n
  • 0.5 to 1 cm tail digested in 500 ul SE buffer\/proteinase K, 55\u00b0C overnight.<\/li>\n
  • Add pre-warmed saturated NaCl solution(6M) 170 ul, gently mix to a final concentration of 1.5 M.<\/li>\n
  • Add 670 ul of chloroform and mix for 30 to 60 minutes by gentle rotation.<\/li>\n
  • Centrifugate at 12000 rpm for five minutes.<\/li>\n
  • Transfer the supernatant to a fresh tube, add one volume of isopropanol at room temperature and gently mix for five minutes.<\/li>\n
  • Pick DNA or centrifugate to collect DNA.<\/li>\n
  • Wash with 70 percent ethanol for one hour or overnight.<\/li>\n
  • Spin and dry.<\/li>\n
  • Dissolve in Tris 10 mM, pH8.5.<\/li>\n<\/ul>\n

    SE Buffer\/Proteinase K<\/h3>\n\n\n\n\n\n\n\n\n
    \u00a0<\/td>\n\n

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

    		\n

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

    		\n

    For 10 ml Mix<\/strong><\/p>\n<\/td>\n<\/tr>\n

    NaCl<\/td>\n75 mM<\/td>\n5 M<\/td>\n150 \u00b5l<\/td>\n<\/tr>\n
    EDTA pH8.0<\/td>\n25 mM<\/td>\n0.5 M<\/td>\n500 \u00b5l<\/td>\n<\/tr>\n
    SDS<\/td>\n1%<\/td>\n10%<\/td>\n1000 \u00b5l<\/td>\n<\/tr>\n
    H2O<\/td>\n\u00a0<\/td>\n\u00a0<\/td>\n8150 \u00b5l<\/td>\n<\/tr>\n
    Proteinase K<\/td>\n200 \u00b5g\/ml<\/td>\n10 mg\/ml<\/td>\n200 \u00b5l<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

    Reference<\/strong><\/p>\n

    R. MGllenbach, PJL Lagoda and C Welter (1989). An Efficient Salt-Chloroform Extraction of DNA from blood and tissues. Trend in Genetics 5(12):391.<\/p>\n<\/div><\/div><\/div><\/div><\/div>","protected":false},"excerpt":{"rendered":"

    0.5 to 1 cm tail digested in 500 ul SE buffer\/proteinase K, 55\u00b0C overnight.Add pre-warmed saturated NaCl solution(6M) 170 ul, gently mix to a final concentration of 1.5 M.Add 670 ul of chloroform and mix for 30 to 60 minutes by gentle rotation.Centrifugate at 12000 rpm for five minutes.Transfer the supernatant to a fresh tube, […]<\/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":"2026-04-26 20:35:37","action":"change-status","newStatus":"draft","terms":[],"taxonomy":""},"_links":{"self":[{"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/186"}],"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=186"}],"version-history":[{"count":1,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/186\/revisions"}],"predecessor-version":[{"id":187,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/186\/revisions\/187"}],"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=186"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}