{"id":197,"date":"2022-05-16T17:29:01","date_gmt":"2022-05-16T17:29:01","guid":{"rendered":"http:\/\/chaletloree.com\/?page_id=197"},"modified":"2025-09-17T15:31:30","modified_gmt":"2025-09-17T15:31:30","slug":"dna","status":"publish","type":"page","link":"http:\/\/chaletloree.com\/index.php\/dna\/","title":{"rendered":"DNA"},"content":{"rendered":"\n<p><u>General techniques<\/u><\/p> <ul> <li><a href=\"https:\/\/docs.google.com\/document\/d\/166DL1bPlqLxL4d0HhMtJAB3NMEo-7EWx7oWS8g5fKtQ\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Procedure for incoming plasmids<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1jc17aVm_N4zRssSpG_quWbwjE661qN78looSfUYYJXk\/edit?usp=sharing\">Preparation of glycerol stocks<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1CZqY-x1KUFCRws7vElU1ZIwz_2hRwuSJ16lj5wkrsP8\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Ethanol precipitation of DNA or RNA<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1jHRM3giyHRMq5RT855YbVjp7m2eQp7Z0VoUwdQwqoSs\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Phenol-Chloroform extraction of nucleic acids<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1GnyC6E0hCxeCRarhVfYQ10OqXV_bBrkm__y2xUIPcbo\/edit?usp=sharing\" style=\"background-color: rgb(255, 255, 255); font-family: rubik, Helvetica, Tahoma, Geneva, Arial, sans-serif; font-size: 1rem;\" title=\"\">Cleavage of DNA into mononucleotides<\/a><\/li> <\/ul> <p><u>Plasmid DNA isolation<\/u><\/p> <ul> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1w3acZNBvCga5mmbkowd6EOFsivN_KSly_-nt5wuL_RQ\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Minipreps using Promega kit <\/a><span style=\"font-family: Helvetica,Arial,sans-serif;\">(also available in <\/span><a href=\"https:\/\/docs.google.com\/document\/d\/1TTXTqfyPOgim4N3uCHeHoQbVKGIlTGAbB7z8cqQIxzY\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">french<\/a><span style=\"font-family: Helvetica,Arial,sans-serif;\">) <\/span><\/li> <li><span style=\"font-family: Helvetica,Arial,sans-serif;\">Midipreps using Macherey &amp; Nagel Kit <\/span><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1aGnk9pqmFyRsmblMyD8EeXk5_QFODgs2PE8w8WnbkPM\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">BAC miniprep<\/a><\/li> <\/ul> <p><u><span style=\"font-family: Helvetica,Arial,sans-serif;\">Plasmid manipulation<\/span><span style=\"font-family: Helvetica,Arial,sans-serif;\"><\/span><\/u><\/p> <ul> <li><span style=\"font-family: Helvetica,Arial,sans-serif;\">Ligation (also available in <\/span><a href=\"https:\/\/docs.google.com\/document\/d\/15uCYtRYLctmb1_Hiw4j0-pYgcSrT8b_yZ5PllDZrAvQ\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">french<\/a><span style=\"font-family: Helvetica,Arial,sans-serif;\">): excel sheet for calculations <\/span><\/li> <li><span style=\"font-family: Helvetica,Arial,sans-serif;\">Gibson assembly: <a href=\"https:\/\/docs.google.com\/spreadsheets\/d\/14t8TrHmI9X4NM03xlDU4HJHfqzw-15kdsWVgRAw7QQ8\/edit?usp=sharing\" title=\"\">excel sheet for calculations <\/a><\/span><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1yMIX8YdAqM5OyQq3H9eJ9DvELa3AZmk4zRG2pKUJ58Q\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Annealing and ligation of linkers <\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1aV7-zzfZvqSHnpR52UzOjcDxy_q4vRnTgZBlPlk3YLs\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Recombination of BAC with PCR product<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1cqtLpQVKgTT6RkWXeC8jEmCyCmYJmltqSXuLW-LbZBQ\/edit\" style=\"font-family: Helvetica,Arial,sans-serif;\">Restriction enzyme digestion<\/a>&nbsp;(also in <a href=\"https:\/\/docs.google.com\/document\/d\/1JmleZ6V8X1CIgeacwlibqYlQ7wG0ylbk9A72ZnD4rbA\/edit?usp=sharing\" title=\"\">french<\/a><a href=\"https:\/\/docs.google.com\/document\/d\/1JmleZ6V8X1CIgeacwlibqYlQ7wG0ylbk9A72ZnD4rbA\/edit?usp=sharing\" title=\"\"> with agarose gel <\/a><a href=\"https:\/\/docs.google.com\/document\/d\/1JmleZ6V8X1CIgeacwlibqYlQ7wG0ylbk9A72ZnD4rbA\/edit?usp=sharing\" title=\"\">elecrophoresis<\/a>)<\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1ff7ds9vtjnIzDk7dwyztK849306oK6NhRaX6faPK3FA\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Dephosphorylation of plasmids<\/a><\/li> <\/ul> <p><u>Transformation and preparation of competent cells<\/u><\/p> <ul> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1GuEC8xbYusMWyUDouZC3ZAUyuEvKlkw-_K88pIOjhmQ\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Preparation of electrocompetent cells<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1vxQFt6b04vm_ykTDs6_r-8XdokJzInw1xIlFRQv-j5o\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Electroporation of electrocompetent cells<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/10dbnxw1Un5P1Juv3gZxwv6yknJG5py-kCPovY8I5vbA\/edit\" title=\"\">Rapid preparation of electrocompetent cells<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1T-zqrGb6iYy2xAfloe3te2i4B-C907ZXpPh4itsJTZA\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Preparation of chemically competent cells<\/a>&nbsp; (also in <a href=\"https:\/\/docs.google.com\/document\/d\/1uOMCoQBwW0a-kmWJufI38-hWx0B1qNmEeK8uHk_fs30\/edit?usp=sharing\" title=\"\">french<\/a>)<\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1o478SWXJVW4qT3h6P_gvlqudh-dXSefl1Mp3aKl-uhI\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Transformation into chemically competent cells<\/a><\/li> <\/ul> <p><u><span style=\"font-family: Helvetica,Arial,sans-serif;\">Extraction of DNA from mammalian cells:<\/span><\/u><\/p> <ul> <li><span style=\"font-family: Helvetica,Arial,sans-serif;\">genomic DNA extraction <\/span> <ul> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1zyoPKziypQALvlw1Ng3ajaaw-ZpR2F4hOhm3B08Vzos\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">12-well plate<\/a><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1twNpyWuRhhNr_nTKu9EJS2IqOhTSElEoOgj-sbZLN0w\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">96-well plate<\/a><\/li> <li><span style=\"font-family: Helvetica,Arial,sans-serif;\">mouse tail <\/span><\/li> <li><a href=\"https:\/\/docs.google.com\/document\/d\/1dZdXdCrBvX7eyANLvyeRaWyFQVjo_ttt0gItMWJw0X0\/edit?usp=sharing\" style=\"font-family: Helvetica,Arial,sans-serif;\">Isolation of DNA from TRIZOL<\/a><\/li> <\/ul> <\/li> <li>extrachromosomal DNA <ul> <li>&nbsp;<\/li> <\/ul> <\/li> <\/ul> <p><br \/> &nbsp;<\/p> <h2 style=\"font-family: Helvetica,Arial,sans-serif; font-size: 11px;\">&nbsp;<\/h2> <h2 style=\"font-family: Helvetica,Arial,sans-serif; font-size: 11px;\">&nbsp;<\/h2> <h2 style=\"font-family: Helvetica,Arial,sans-serif;\">&nbsp;<\/h2> \n\n","protected":false},"excerpt":{"rendered":"<p>General techniques Procedure for incoming plasmids Preparation of glycerol stocks Ethanol precipitation of DNA or RNA Phenol-Chloroform extraction of nucleic acids Cleavage of DNA into mononucleotides Plasmid DNA isolation Minipreps using Promega kit (also available in french) Midipreps using Macherey &amp; Nagel Kit BAC miniprep Plasmid manipulation Ligation (also available in french): excel sheet for [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-197","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/197","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/comments?post=197"}],"version-history":[{"count":3,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/197\/revisions"}],"predecessor-version":[{"id":293,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/197\/revisions\/293"}],"wp:attachment":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/media?parent=197"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}