{"id":25322,"date":"2025-12-14T09:01:56","date_gmt":"2025-12-14T00:01:56","guid":{"rendered":"https:\/\/blog.agentsoft.co.kr\/index.php\/2025\/12\/14\/25322\/"},"modified":"2025-12-14T09:01:56","modified_gmt":"2025-12-14T00:01:56","slug":"%ed%94%8c%eb%9d%bc%ec%8a%a4%eb%af%b8%eb%93%9c-dna-%ec%a0%95%ec%a0%9c-purification-of-plasmid-dna","status":"publish","type":"post","link":"https:\/\/blog.agentsoft.co.kr\/index.php\/2025\/12\/14\/25322\/","title":{"rendered":"\ud50c\ub77c\uc2a4\ubbf8\ub4dc DNA \uc815\uc81c\/ purification of plasmid DNA"},"content":{"rendered":"

<\/p>\n

\ubaa9\ucc28<\/strong><\/p>\n

1. Summary
\n2. Introduction
\n3. Material and Methods
\n4. Result and Discussion
\n5. Reference<\/p>\n

\ubcf8\ubb38\ub0b4\uc6a9<\/strong><\/p>\n

1) Plasmid DNA \ubd84\ub9ac <\/p>\n

Plasmid DNA\ub294 \uc7ac\uc870\ud569 DNA \uc870\uc791\uc744 \uc704\ud55c \uc6b4\ubc18\uccb4 DNA\ub85c \uc0ac\uc6a9\ub418\ub294\ub370, \uc774 \uc6b4\ubc18\uccb4 DNA\ub97c Vector\ub77c\uace0 \ud55c\ub2e4. \uc8fc\ub85c \ub300\uc7a5\uade0\uc5d0 vector\ub97c \uc0bd\uc785\ud558\ub294\ub370 \uc774 \uacfc\uc815\uc744 Bacterial Transformation\uc774\ub77c\uace0 \ud55c\ub2e4. vector\ub97c cell\uc5d0 \ub3c4\uc785\ud55c \ub4a4, \uc120\ubcc4\uc744 \uc704\ud574 ampicillin\uc774 \ud568\uc720\ub41c \ubc30\uc9c0\uc5d0\uc11c \ud0a4\uc6b0\ub294 \uacfc\uc815\uc744 \uc9c4\ud589\ud55c\ub2e4. ampicillin\uc774 \ud568\uc720\ub41c \ubc30\uc9c0\uc5d0\uc11c \uc790\ub77c\ub09c colony\ub294 Bacterial Transformation\uc774 \uc131\uacf5\ud55c cell\uc774\ub2e4. \ub300\uc7a5\uade0 cell\uc5d0\ub294 plasmid DNA\uc640 \ub300\uc7a5\uade0 cell\uc758 \uc5fc\uc0c9\uccb4 DNA\uc778 chromosomal DNA\uac00 \ub3c5\uc790\uc801\uc73c\ub85c \uc874\uc7ac\ud558\uae30 \ub54c\ubb38\uc5d0 plasmid DNA\ub97c \ub530\ub85c \ubd84\ub9ac\ud558\uc5ec\uc57c \ud55c\ub2e4. cell\uc5d0\uc11c plasmid DNA\ub97c \ub530\ub85c \ubd84\ub9ac\ud558\ub294 \uacfc\uc815\uc744 Purification of plasmid DNA\ub77c\uace0 \ud55c\ub2e4. plasmid DNA \ubd84\ub9ac\uc5d0\ub294 boiling method, alkaline lysis, lithium based procedure \ub4f1 \uc5ec\ub7ec \ubc29\ubc95\uc774 \uc788\uc9c0\ub9cc \uac00\uc7a5 \ubcf4\ud3b8\uc801\uc73c\ub85c \uc0ac\uc6a9\ub418\ub294 mini-prep \ubc29\ubc95\uc740 alkaline lysis\uc774\ub2e4. \ub192\uc740 pH\uc758 \uc54c\uce7c\ub9ac\uc131 \uc6a9\uc561\uc744 \uc0ac\uc6a9\ud558\uc5ec cell\uc758 \uc138\ud3ec\ubcbd\uacfc \uc138\ud3ec\ub9c9\uc744 \uc6a9\ud574\uc2dc\ud0a8 \ud6c4, \uc0b0\uc131 \uc6a9\uc561\uc744 \ucc98\ub9ac\ud558\uc5ec plasmid DNA\ub9cc\uc744 \ubd84\ub9ac\ud574\ub0b4\ub294 \ubc29\ubc95\uc774\ub2e4. \uc54c\uce7c\ub9ac\uc131 \uc6a9\uc561\uc774 cell\uc744 \uc6a9\ud574\uc2dc\ud0a4\uba74 cell \uc548\uc758 chromosomal DNA\uc640 \uc218\ud655\ud574\uc57c \ud560 plasmid DNA\uac00 \ub178\ucd9c\ub418\uc5b4 \ubcc0\uc131\uc774 \uc77c\uc5b4\ub098\uac8c \ub41c\ub2e4. \ub450 DNA \ubaa8\ub450 \ubcc0\uc131\uc774 \uc77c\uc5b4\ub098\uc9c0\ub9cc \uc0b0\uc131 \uc6a9\uc561\uc744 \uc774\uc6a9\ud558\uc5ec \uc6a9\uc561\uc744 \uc911\uc131\uc73c\ub85c \ub9cc\ub4e4\uc5b4\uc8fc\uba74 \uc6d0\ub798\uc758 \ubcc0\uc131 \uc804 \uc0c1\ud0dc\ub85c \ubcf5\uc6d0\uc774 \ub41c\ub2e4. \ud558\uc9c0\ub9cc chromosomal DNA\ub294 \ud06c\uae30\uac00 \ud06c\uace0 \uad6c\uc870\uac00 \ubcf5\uc7a1\ud558\uc5ec \ubcc0\uc131 \uc804\uc758 \uc0c1\ud0dc\ub85c \ubcf5\uc6d0\uc774 \ub418\uc9c0 \ubabb\ud558\uace0 \uce68\uc804\uc774 \ub418\uc5b4 \uc81c\uac70\ub418\uba70, plasmid DNA\ub294 \ube44\uad50\uc801 \uac04\ub2e8\ud55c \uad6c\uc870\uc774\uae30 \ub54c\ubb38\uc5d0 \uc6d0\ub798 \uc0c1\ud0dc\ub85c \ubcf5\uc6d0\ub418\uac8c \ub41c\ub2e4. \ubd84\ub9ac\uac00 \ub05d\ub09c plasmid DNA\ub294 Electrophoresis\ub97c \ud1b5\ud574 \ud655\uc778\ud560 \uc218 \uc788\ub2e4. <\/p>\n

2) Alkali lysis<\/p>\n

Alkali lysis\ub294 mini-prep\uc758 \ud55c \uc885\ub958\ub85c \uc6a9\uc561\uc758 pH\ub97c \uc774\uc6a9\ud558\uc5ec plasmid DNA\ub97c \ubd84\ub9ac\ud558\ub294 \ubc29\ubc95\uc774\ub2e4. \uc5ec\uae30\uc11c mini-prep\uc740 DNA purification\uc758 \ud55c \uc885\ub958\ub85c mini\ub294 DNA\ub97c \uc18c\ub7c9 \ucd94\ucd9c\ud55c\ub2e4\ub294 \ub73b\uc774\uba70 prep\uc740 preparation\uc744 \uc758\ubbf8\ud55c\ub2e4. Alkali lysis\ub294 Re-suspension, Cell lysis, Neutralization, Cleaning and Concentration \uc21c\uc11c\ub85c \uc9c4\ud589\ud558\uac8c \ub41c\ub2e4. Re-suspension \ub2e8\uacc4\uc5d0\uc11c\ub294 S1 buffer\ub97c \uc0ac\uc6a9\ud558\uac8c \ub418\ub294\ub370, \uc774 buffer\ub294 glucose, EDTA(chelator), RNase, Tris \ub4f1\uc774 \ud568\uc720\ub418\uc5b4 \uc788\ub2e4.<\/p>\n

<\uc911 \ub7b5><\/p>\n

Nanodrop \ubd84\uc11d \uacb0\uacfc 4\uc870\ub294 1\ubc88 plasmid DNA\ub294 \ub18d\ub3c4\uac00 \ub2e4\ub978 \uc870\uc5d0 \ube44\ud574 \uaf64 \ub192\uc740 \ud3b8\uc778 206.3 ng\/\u00b5L\uac00 \ub098\uc654\uace0 2\ubc88 plasmid\ub294 \ub2e4\ub978 \ud300\uc5d0 \ube44\ud574 DNA \ub18d\ub3c4\uac00 \ub0ae\uc740 \ud3b8\uc778 38.9 ng\/\u00b5L\uac00 \ub098\uc654\ub2e4.
\nA260\/A280 \ube44\uc728\uc758 \uc218\uce58\ub294 \ub2e8\ubc31\uc9c8\uc5d0 \ub300\ud55c \ud575\uc0b0\uc758 \uc591\uc744 \ub098\ud0c0\ub0b8\ub2e4. \uc21c\uc218\ud558\uac8c \uc815\uc81c\ub41c RNA\ub098 DNA\ub294 \uac01\uac01 2.1\uacfc 1.8\uc758 \uc218\uce58\ub97c \ub098\ud0c0\ub0b8\ub2e4. \ube44\uc728\uc774 \ub0ae\uc744\uc218\ub85d \ub2e8\ubc31\uc9c8\uc758 \ud568\ub7c9\uc774 \ub192\ub2e4\ub294 \uc758\ubbf8\uc774\uba70 \ub192\uc740 \ub2e8\ubc31\uc9c8 \ud568\ub7c9\uc740 \ud575\uc0b0\uc744 \uc774\uc6a9\ud558\ub294 \uc2e4\ud5d8\uc5d0 \uc601\ud5a5\uc744 \ubbf8\uce60 \uc218 \uc788\ub2e4. \ubaa8\ub4e0 \uc870\uc5d0\uc11c A260\/A280 \ube44\uc728\uc740 1.8\ubcf4\ub2e4 \ub192\uae30 \ub54c\ubb38\uc5d0 \ub2e8\ubc31\uc9c8 \ud568\ub7c9\uc774 \ub192\uc9c0 \uc54a\uac8c \ubd84\ub9ac\uac00 \uc798 \ub418\uc5c8\ub2e4\ub294 \uac83\uc744 \uc54c \uc218 \uc788\ub2e4.<\/p>\n

\ucd9c\ucc98 : \ud574\ud53c\ucea0\ud37c\uc2a4<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"

\ubaa9\ucc28 1. Summary 2. Introduction 3. Material and Methods 4. Result and Discussion 5. Reference \ubcf8\ubb38\ub0b4\uc6a9 1) Plasmid DNA \ubd84\ub9ac Plasmid DNA\ub294 \uc7ac\uc870\ud569 DNA \uc870\uc791\uc744 \uc704\ud55c \uc6b4\ubc18\uccb4 DNA\ub85c \uc0ac\uc6a9\ub418\ub294\ub370, \uc774 \uc6b4\ubc18\uccb4 DNA\ub97c Vector\ub77c\uace0 \ud55c\ub2e4. \uc8fc\ub85c \ub300\uc7a5\uade0\uc5d0 vector\ub97c \uc0bd\uc785\ud558\ub294\ub370 \uc774 \uacfc\uc815\uc744 Bacterial Transformation\uc774\ub77c\uace0 \ud55c\ub2e4. vector\ub97c cell\uc5d0 \ub3c4\uc785\ud55c \ub4a4, \uc120\ubcc4\uc744 \uc704\ud574 ampicillin\uc774 \ud568\uc720\ub41c \ubc30\uc9c0\uc5d0\uc11c \ud0a4\uc6b0\ub294 \uacfc\uc815\uc744 \uc9c4\ud589\ud55c\ub2e4. […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[31506,31505,10165,8835,31504],"class_list":["post-25322","post","type-post","status-publish","format-standard","hentry","category-uncategorized","tag-dna-purification","tag-dna-","tag-plasmid-dna","tag-8835","tag--dna"],"_links":{"self":[{"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/posts\/25322","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/comments?post=25322"}],"version-history":[{"count":0,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/posts\/25322\/revisions"}],"wp:attachment":[{"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/media?parent=25322"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/categories?post=25322"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/tags?post=25322"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}