{"id":171,"date":"2017-04-19T15:21:15","date_gmt":"2017-04-19T19:21:15","guid":{"rendered":"https:\/\/health.uconn.edu\/mouse-genome-modification\/?page_id=171"},"modified":"2017-04-19T15:21:15","modified_gmt":"2017-04-19T19:21:15","slug":"dna-preparation-for-pcr-product-less-than-500-bp","status":"publish","type":"page","link":"https:\/\/health.uconn.edu\/mouse-genome-modification\/protocols\/dna-preparation-for-pcr-product-less-than-500-bp\/","title":{"rendered":"DNA Preparation for PCR Product Less Than 500 Bp"},"content":{"rendered":"

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

\u00a0<\/strong><\/p>\n<\/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

Tris pH8.8<\/td>\n50 mM<\/td>\n1 M<\/td>\n500 \u00b5l<\/td>\n<\/tr>\n
EDTA pH 8.0<\/td>\n1 mM<\/td>\n0.5 M<\/td>\n20 \u00b5l<\/td>\n<\/tr>\n
Tween<\/td>\n0.5%<\/td>\n10%<\/td>\n500 \u00b5l<\/td>\n<\/tr>\n
H2O<\/td>\n\u00a0<\/td>\n\u00a0<\/td>\n9000 \u00b5l<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
    \n
  • Add Proteinase K before use.<\/li>\n
  • For tail piece 0.2 mm long or ear piece, add 50 ul buffer plus 1.5 ul of 20 mg\/ml (30 ug) Proteinase K stock solution.<\/li>\n
  • Ten samples: 550 ul lysis buffer + 16.5 ul PK 20 mg\/ml.<\/li>\n
  • You can use 0.5 ml PCR tubes for digestion and heat in a PCR machine.<\/li>\n<\/ul>\n

    Lysis<\/h2>\n
      \n
    • 55\u00b0C, O\/N , then heat inactive at 100\u00b0C for ten minutes. Cool down on bench top.<\/li>\n
    • For 20 ul PCR reaction, use 0.8 ul tail DNA.<\/li>\n<\/ul>\n<\/div><\/div><\/div><\/div><\/div>","protected":false},"excerpt":{"rendered":"

      Lysis Buffer\u00a0Final ConcentrationStock ConcentrationFor 10 ml MixTris pH8.850 mM1 M500 \u00b5lEDTA pH 8.01 mM0.5 M20 \u00b5lTween0.5%10%500 \u00b5lH2O\u00a0\u00a09000 \u00b5lAdd Proteinase K before use.For tail piece 0.2 mm long or ear piece, add 50 ul buffer plus 1.5 ul of 20 mg\/ml (30 ug) Proteinase K stock solution.Ten samples: 550 ul lysis buffer + 16.5 ul PK […]<\/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":"2026-04-10 23:12:27","action":"change-status","newStatus":"draft","terms":[],"taxonomy":""},"_links":{"self":[{"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/171"}],"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=171"}],"version-history":[{"count":1,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/171\/revisions"}],"predecessor-version":[{"id":172,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/171\/revisions\/172"}],"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=171"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}