{"id":1462,"date":"2025-04-20T09:01:03","date_gmt":"2025-04-20T00:01:03","guid":{"rendered":"https:\/\/blog.agentsoft.co.kr\/index.php\/2025\/04\/20\/1462\/"},"modified":"2025-04-20T09:01:03","modified_gmt":"2025-04-20T00:01:03","slug":"%eb%af%b8%ec%83%9d%eb%ac%bc-%eb%a9%b4%ec%97%ad%ed%95%99-%ec%8b%a4%ed%97%98%eb%b3%b4%ea%b3%a0%ec%84%9c","status":"publish","type":"post","link":"https:\/\/blog.agentsoft.co.kr\/index.php\/2025\/04\/20\/1462\/","title":{"rendered":"\ubbf8\uc0dd\ubb3c \uba74\uc5ed\ud559 \uc2e4\ud5d8\ubcf4\uace0\uc11c"},"content":{"rendered":"<p><img decoding=\"async\" src=\"https:\/\/image4.happycampus.com\/Production\/thumb212\/2024\/01\/03\/data29316497-0001.jpg\"><img decoding=\"async\" src=\"https:\/\/image4.happycampus.com\/Production\/thumb212\/2024\/01\/03\/data29316497-0002.jpg\"><\/p>\n<p><strong>\ubaa9\ucc28<\/strong><\/p>\n<p>1. Preparation of media (LA\/LB &amp; TGY broth-agar)<br \/>\n2. Bacteria cell culture<br \/>\n3. Genome prep (Genomic DNA extraction)<br \/>\n4. PCR for 16S rRNA sequencing<br \/>\n5. Agarose gel Electrophoresis<br \/>\n6. DNA sequencing analysis<br \/>\n7. Measurement of CFU<br \/>\n8. Isolation of non-pigment colonies<br \/>\n9. PCR for carotenoid genes<br \/>\n10. Plasmid mini prep<br \/>\n11. Transformation<br \/>\n12. Data analysis<\/p>\n<p><strong>\ubcf8\ubb38\ub0b4\uc6a9<\/strong><\/p>\n<p>-\uc2e4\ud5d8\uacfc\uc815<br \/>\n-Cell harvesting for Gram-Positive bacteria<br \/>\n1. 70\u00b0C incubator\ub97c \ubbf8\ub9ac \ucf1c\uc11c \uc628\ub3c4\ub97c \uc62c\ub824 \ub193\ub294\ub2e4.<br \/>\n2. Lysozyme buffer (sample \ub2f9 0.004g lysozyme + 200\u03bcl (\ub9c8\uc774\ud06c\ub85c\ub9ac\ud130; \u03bb = \ub78c\ub2e4) GPT buffer) \ub97c \uc900\ube44\ud55c\ub2e4.<br \/>\n3. \uc720\uc804\uccb4\ub97c \ubf51\uace0\uc790 \ud558\ub294 \uade0\uc744 overnight-culture(12~16h)\uc2dc\ud0a8 \ud6c4, EP-tube\uc5d0 1.5ml\uc529 \ub2f4\uace0 2\ubd84\uac04 centrifuge \uc2dc\ud0a8\ub2e4. \uadf8 \ud6c4, \uc0c1\uce35\uc561\uc744 \ubaa8\ub450 \ubc84\ub9b0\ub2e4.<br \/>\n4. Lysozyme buffer 200\u03bcl\ub97c \ub123\uace0, pipetting\uc73c\ub85c \uc798 \uc11e\uc5b4\uc900\ub2e4.<br \/>\n5. 10\ubd84\uac04 \uc2e4\ub0b4\uc628\ub3c4\ub85c incubation\uc2dc\ud0a8\ub2e4. (2~3\ubd84\ub9c8\ub2e4 \uac00\ubccd\uac8c \ud754\ub4e4\uc5b4\uc904 \uac83)<br \/>\n-Cell lysis : Lysis buffer\ub294 \uc138\ud3ec\ub9c9\uc744 \uae68\ub294 \uc6a9\ub3c4<br \/>\n6. GB buffer 200\u03bcl\ub97c \ub123\uace0, 5\ucd08\uac04 pipetting \ud574\uc900\ub2e4.<br \/>\n7. 70\u00b0C incubator\uc5d0 \uc900\ube44\ud55c sample\uc744 \ub123\uace0 10\ubd84\uac04 incubation\uc2dc\ud0a8\ub2e4. (3\ubd84\ub9c8\ub2e4 \uac00\ubccd\uac8c \ud754\ub4e4\uc5b4\uc904 \uac83) \uc774\ub54c, D.W\ub97c 70\u00b0C incubator\uc5d0 \ub123\uc5b4\ub454\ub2e4.(elution) \uadf8\ub9ac\uace0 Collection tube\uc5d0 GB Column\uc744 \ubbf8\ub9ac \ub07c\uc6cc \ub123\uace0, labeling\ud55c\ub2e4.<br \/>\n-DNA Binding<br \/>\n8. \uc5d0\ud0c4\uc62c (96~100%) 200\u03bcl\ub97c \ub123\uace0, \uc989\uc2dc vortexing\uc73c\ub85c 10\ucd08\uac04 \uc11e\uc5b4\uc900\ub2e4.<br \/>\n9. EP-tube\uc5d0 \ub2f4\uae34 sample\uc744 \uc900\ube44\ud55c GB Column\uc5d0 \ub123\ub294\ub2e4.<br \/>\n10. cap\uc744 \ub2eb\uace0 2\ubd84 30\ucd08\uac04 centrifuge (14,000-16,000 x g) \uc2dc\ud0a8\ub2e4. \uadf8 \ud6c4, \uac78\ub7ec\uc9c4 \uc561\uccb4\ub97c \ubc84\ub9b0\ub2e4.<br \/>\n-Wash : Column\uacfc Bead\uc5d0 \ubd80\ucc29\ub41c DNA \ub5a8\uc5b4\ub728\ub9ac\uae30<br \/>\n11. W1 Buffer 400\u03bcl\ub97c GB Column\uc5d0 \ub123\uc740 \ud6c4, 2\ubd84\uac04 centrifuge (14,000-16,000 x g) \uc2dc\ud0a4\uace0 \uac78\ub7ec \uc9c4 \uc561\uccb4\ub97c \ubc84\ub9b0\ub2e4.<br \/>\n12. (\uc5d0\ud0c4\uc62c\uc774 \ucca8\uac00\ub41c) Wash Buffer 600\u03bcl\ub97c GB Column\uc5d0 \ub123\ub294\ub2e4. 2\ubd84\uac04 centrifuge (14,000- 16,000 x g) \uc2dc\ud0a4\uace0 \uac78\ub7ec\uc9c4 \uc561\uccb4\ub97c \ubc84\ub9b0\ub2e4.<\/p>\n<p>\ucd9c\ucc98 : <a href=\"https:\/\/www.happycampus.com\/report-doc\/29316497\/\" target=\"_blank\">\ud574\ud53c\ucea0\ud37c\uc2a4<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>\ubaa9\ucc28 1. Preparation of media (LA\/LB &amp; TGY broth-agar) 2. Bacteria cell culture 3. Genome prep (Genomic DNA extraction) 4. PCR for 16S rRNA sequencing 5. Agarose gel Electrophoresis 6. DNA sequencing analysis 7. Measurement of CFU 8. Isolation of non-pigment colonies 9. PCR for carotenoid genes 10. Plasmid mini prep 11. Transformation 12. Data [&hellip;]<\/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":[2631,2633,1386,2629,261,2630,2622,2632],"class_list":["post-1462","post","type-post","status-publish","format-standard","hentry","category-uncategorized","tag-genome-prep","tag-pcr","tag-1386","tag-2629","tag-261","tag-2630","tag-2622","tag-2632"],"_links":{"self":[{"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/posts\/1462","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=1462"}],"version-history":[{"count":0,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/posts\/1462\/revisions"}],"wp:attachment":[{"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/media?parent=1462"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/categories?post=1462"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/blog.agentsoft.co.kr\/index.php\/wp-json\/wp\/v2\/tags?post=1462"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}