{"id":179,"date":"2017-04-19T15:24:55","date_gmt":"2017-04-19T19:24:55","guid":{"rendered":"https:\/\/health.uconn.edu\/mouse-genome-modification\/?page_id=179"},"modified":"2025-08-14T14:10:05","modified_gmt":"2025-08-14T18:10:05","slug":"allele-validation","status":"publish","type":"page","link":"https:\/\/health.uconn.edu\/mouse-genome-modification\/protocols\/allele-validation\/","title":{"rendered":"Allele Validation for Genetically Modified F1 Mice"},"content":{"rendered":"

PCR Conditions for Genotyping<\/h2>\n

You will receive primer sequences for the gene targeting mice. The following conditions are used for PCR genotyping for all our gene targeting mice. We'll email you if special conditions are used for your mice.<\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

PCR Mix - Stocks<\/strong><\/p>\n<\/td>\n

\n

One Reaction<\/strong><\/p>\n<\/td>\n

\n

Ten Reactions (Make for 11 Reactions)<\/strong><\/p>\n<\/td>\n<\/tr>\n

dNTP 10mM<\/td>\n0.25 \u00b5l<\/td>\n2.75 \u00b5l<\/td>\n<\/tr>\n
10x PCR Buffer<\/td>\n1.25 \u00b5l<\/td>\n13.75 \u00b5l<\/td>\n<\/tr>\n
Primers 10 uM<\/td>\n0.3 \u00b5l each<\/td>\n3.3 \u00b5l each<\/td>\n<\/tr>\n
Taq Polymerase<\/td>\n0.06 \u00b5l<\/td>\n0.66 \u00b5l<\/td>\n<\/tr>\n
H2O<\/td>\n9.48 \u00b5l<\/td>\n104.28 \u00b5l<\/td>\n<\/tr>\n
Total Volume<\/td>\n12.02 \u00b5l<\/td>\n124.74 \u00b5l<\/td>\n<\/tr>\n
Add DNA<\/a>: 0.5 \u00b5l<\/td>\n\u00a0<\/td>\n\u00a0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

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

PCR Cycles<\/strong><\/p>\n<\/td>\n

\n
\u00a0<\/h5>\n<\/td>\n
\n
\u00a0<\/h5>\n<\/td>\n<\/tr>\n
1. Denaturation:<\/td>\n94\u00b0C<\/td>\n3 minutes<\/td>\n<\/tr>\n
2. Denaturation:<\/td>\n94\u00b0C<\/td>\n30 seconds<\/td>\n<\/tr>\n
3. Annealing:<\/td>\n55 \u00b0C<\/td>\n30 seconds<\/td>\n<\/tr>\n
4. Extension: 2-4 steps X 35 cycles<\/td>\n72 \u00b0C<\/td>\n30 seconds<\/td>\n<\/tr>\n
5. Extension<\/td>\n72\u00b0C<\/td>\n5 minutes<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div><\/div><\/div><\/div><\/div>","protected":false},"excerpt":{"rendered":"

PCR Conditions for GenotypingYou will receive primer sequences for the gene targeting mice. The following conditions are used for PCR genotyping for all our gene targeting mice. We’ll email you if special conditions are used for your mice.PCR Mix – StocksOne ReactionTen Reactions (Make for 11 Reactions)dNTP 10mM0.25 \u00b5l2.75 \u00b5l10x PCR Buffer1.25 \u00b5l13.75 \u00b5lPrimers 10 […]<\/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\/179"}],"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=179"}],"version-history":[{"count":3,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/179\/revisions"}],"predecessor-version":[{"id":1223,"href":"https:\/\/health.uconn.edu\/mouse-genome-modification\/wp-json\/wp\/v2\/pages\/179\/revisions\/1223"}],"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=179"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}