{"id":44527,"date":"2023-07-28T13:50:17","date_gmt":"2023-07-28T05:50:17","guid":{"rendered":"https:\/\/biopioneer.com.tw\/?p=44527"},"modified":"2023-11-09T17:41:58","modified_gmt":"2023-11-09T09:41:58","slug":"proclone-%e9%ab%98%e6%95%88%e8%83%bdcompetent-cell%e5%8b%9d%e4%bb%bb%e7%b4%b0%e8%83%9e-abmgood%e8%b2%a8%e8%99%9fe003","status":"publish","type":"post","link":"https:\/\/biopioneer.com.tw\/?p=44527","title":{"rendered":"ProClone\u2122 \u9ad8\u6548\u80fdCompetent cell\u52dd\u4efb\u7d30\u80de | ABMGOOD\u8ca8\u865fE003"},"content":{"rendered":"

ProClone\u2122 \u9ad8\u6548\u80fdCompetent cell\u52dd\u4efb\u7d30\u80de | ABMGOOD\u8ca8\u865fE003<\/span><\/strong><\/a><\/span><\/p>\n

\u5177\u5099\u6709\u9ad8\u6548\u80fd\u7684Competent cell\u52dd\u4efb\u7d30\u80de\uff0c\u662fcloning\u6280\u8853\u57fa\u672c\u8981\u9818\u3002ProClone\u2122<\/strong>\u8b93Competent cell\u63da\u540d\u5168\u4e16\u754c\u3002\u5176\u9ad8\u6548\u80fd\u7684\u80fd\u529b\uff0c\u8b93cloning\u66f4\u5bb9\u6613\uff0c\u5728\u4e00\u5206\u9418\u5c31\u505a\u5b8cTransformation\u3002\u8f49\u5316\u6548\u7387\uff1a>1 x 10^6 CFU\/\u03bcg DNA\u3002\u7522\u751f\u9ad8\u8cea\u91cf\u7684\u8cea\u7c92\u88fd\u5291 (endA1)\u3001\u6297 T1 \u566c\u83cc\u9ad4 (fhuA2)\u3001\u91cd\u7d44\u7387\u964d\u4f4e (recA1)\u3001\u85cd\/\u767d\u7be9\u9078\u80fd\u529b\u3002<\/p>\n

Features:<\/span><\/strong><\/span><\/p>\n

Transformation efficiency: >1 x 10^6 CFU\/\u00b5g of DNA
\nResults in high quality plasmid preparations (endA1)
\nResistant to T1 phage (fhuA2)
\nExhibits reduced recombination rate (recA1)
\nBlue\/White screening capable<\/p>\n

<\/p>\n\n\n\n\n\n\n\n\n\n
Cat. No.<\/td>\nE003<\/span><\/td>\n<\/tr>\n
Name<\/td>\nProClone\u2122 Competent Cells<\/td>\n<\/tr>\n
Unit<\/td>\n5ml (4 x 1.25ml)<\/td>\n<\/tr>\n
Category<\/td>\nMolecular Biology Enzymes and Kits<\/td>\n<\/tr>\n
Description<\/td>\nProClone\u2122 Competent Cells are high-efficiency, chemically competent DH5\u03b1 (E. coli<\/em>) cells, ideal for both routine and challenging plasmid amplification and subcloning projects.<\/td>\n<\/tr>\n
Expression System General<\/td>\nE. coli<\/td>\n<\/tr>\n
Storage Condition<\/td>\nStore at -80\u00b0C.<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Protocol <\/span><\/strong><\/p>\n

The following protocol serves as a general guideline and may require optimization.<\/p>\n

1. Thaw 1 tube of cells on ice for 10-15 minutes, then gently invert tube to thoroughly mix any settled cells. Aliquot 60 \u00b5l of cells into a sterile 1.5 ml tube per transformation. Remaining cells can be aliquoted and re-frozen for later use, but expect lower performance from re-frozen cells.<\/p>\n

2. Add 10 pg-100 ng of plasmid DNA or 5-10 \u00b5l of the subcloning reaction (i.e. ligation) to the cells and mix gently. Incubate the mixture at 4\u00b0C for 30 minutes.<\/p>\n

3. Heat shock the cells using a waterbath or heat block at 42\u00b0C for 45 seconds, then immediately place on ice for 1-2 minutes.<\/p>\n

4. Add 150 \u00b5l of sterile LB broth containing no antibiotics and incubate the cells at 37\u00b0C in a shaking incubator for 1 hour for recovery.<\/p>\n

5. Plate the entire volume on LB agar containing appropriate antibiotics and incubate overnight at 37\u00b0C.<\/p>\n

\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df<\/a><\/p>\n

\u3010Competent cell\u52dd\u4efb\u7d30\u80de\u6ce8\u610f\u4e8b\u9805\u3011<\/strong><\/span><\/p>\n

1.\u672c\u5efa\u8b70\u6fc3\u5ea6\u4ee5\u65b0\u9bae\u914d\u88fd\u4e4b\u6297\u751f\u7d20\u70ba\u6e96\u3002<\/p>\n

2.\u4f7f\u7528\u904e\u671f\u4e4b\u6297\u751f\u7d20\uff0c\u5c07\u56e0\u85e5\u6027\u4e0d\u8db3\uff0c\u6613\u7522\u751f\u5047\u6027\u8f49\u5f62\u83cc\u843d\u3002<\/p>\n

3.\u82e5\u4f7f\u7528\u904e\u91cf\u7684\u6297\u751f\u7d20\u6fc3\u5ea6\u6216\u540c\u6642\u4f7f\u7528\u591a\u7a2e\u6297\u751f\u7d20\u9032\u884c\u7be9\u9078\uff0c\u8f49\u5f62\u6548\u7387\u6703\u660e\u986f\u4e0b\u964d\u3002\u8acb\u52d9\u5fc5\u4f9d\u5efa\u8b70\u6fc3\u5ea6\u9032\u884c\u6297\u751f\u7d20\u7be9\u9078<\/p>\n

\u3010\u85cd<\/strong>\/<\/strong>\u767d\u7be9\u9078\u3011<\/strong><\/span><\/p>\n

\u9032\u884c\u8f49\u5f62\u524d\uff0c\u8acb\u5148\u78ba\u8a8d\u8cea\u9ad4\u5e36\u6709Lac<\/em>Z operon\u3002\u8f49\u5f62\u5f8c\uff0c\u8acb\u5c07\u52dd\u4efb\u7d30\u80de\u5857\u4f48\u65bc\u542b0.5 mM IPTG<\/a><\/span> \u548c 40~60 \u03bcg\/ml X-gal\u4e4bLB\u57f9\u990a\u57fa\u300237oC<\/sup>\u57f9\u990a\u5f8c\uff0c\u767d\u8272\u83cc\u843d\u8868\u793a\u5916\u6e90DNA\u5d4c\u5165Lac<\/em>Z operon\uff0c\u85cd\u8272\u83cc\u843d\u5247\u4ecd\u5177\u5099\u6b63\u5e38\u7684\u03b2-galactosidase\u4ee3\u8b1d\u80fd\u529b\u3002<\/p>\n

\u3010\u542b\u6297\u751f\u7d20\u4e4bLB\u57f9\u990a\u57fa\u914d\u65b9\u3011<\/span><\/strong><\/span><\/p>\n

10 g\/L Tryptone 5 g\/L Yeast extract 10 g\/L NaCl 15 g\/L Agar \u4ee5NaOH\u8abf\u6574\u9178\u9e7c\u503c\u81f3pH7.5\uff0c\u9ad8\u6eab\u9ad8\u58d3\u6ec5\u83cc\u5f8c\uff0c\u5f85\u964d\u6eab\u81f355oC\uff0c\u52a0\u5165\u9069\u7576\u6fc3\u5ea6\u4e4b\u6297\u751f\u7d20\u3002<\/p>\n

LB Agar (Miller\u914d\u65b9)\u57f9\u990a\u57fa | \u8ca8\u865fNCM0142A<\/span><\/strong><\/a><\/span><\/p>\n

LB<\/strong>\u57f9\u990a\u57fa<\/strong>\uff08Lysogeny broth<\/strong>\uff0cLB\uff09\u5ee3\u6cdb\u89e3\u91cb\u7232Luria-Bertani\u57f9\u990a\u57fa\uff0c\u4f9d\u64da\u5176\u767c\u660e\u4eba\u8c9d\u723e\u5854\u5c3c\uff08Giuseppe Bertani\uff09 \u7684\u8aaa\u6cd5\uff0c\u9019\u500b\u540d\u5b57\u4f86\u6e90\u65bc\u82f1\u8a9e\u7684 Lysogeny broth\uff0c\u5373\u6eb6\u83cc\u57f9\u990a\u57fa\u3002Luria-Bertani\u57f9\u990a\u57fa\u662f\u5fae\u751f\u7269\u5b78\u5be6\u9a57\u4e2d\u6700\u5e38\u7528\u7684\u71df\u990a\u6027\u57f9\u990a\u57fa\uff0c\u7528\u65bc\u57f9\u990a\u5927\u8178\u687f\u83cc(E. coli<\/em>\u00a0i) \u7b49\u7d30\u83cc\uff0cBroth\u52a0\u5165\u74ca\u8102(Agar)\u88fd\u6210\u7684\u56fa\u614b\u57f9\u990a\u57fa\u3002\u52a0\u5165\u6297\u751f\u7d20\u7684 LB \u57f9\u990a\u57fa(LB Broth) \u53ef\u7528\u65bc\u7be9\u9078\u5927\u8178\u687f\u83cc\u7232\u5bbf\u4e3b\u7684cloning\u3002<\/p>\n

LB Agar\uff08Miller\uff09\u662f\u4e00\u7a2e\u71df\u990a\u8c50\u5bcc\u7684\u57f9\u990a\u57fa\uff0c\u8a2d\u8a08\u7528\u65bc\u57f9\u990a\u91cd\u7d44\u83cc(recombinant strains) \u7684\u7d14\u57f9\u990a\u7269\uff0c\u5b83\u57fa\u65bcMiller\u63cf\u8ff0\u7684\u914d\u65b9\u3002\u5927\u8178\u687f\u83cc\u5728LB\u57f9\u990a\u57fa\u4e2d\u751f\u9577\u81f3\u5c0d\u6578\u665a\u671f (late log phase)\u3002 \u4e00\u4e9bplasmid vectors\u8907\u88fd\u5230high copy number\u4e26\u4e14\u4e0d\u9700\u8981\u9078\u64c7\u6027\u64f4\u589e\u3002 \u4e00\u4e9b\u8f09\u9ad4\u4e0d\u80fd\u5982\u6b64\u81ea\u7531\u5730\u8907\u5236\u4e26\u4e14\u9700\u8981\u88ab\u9078\u64c7\u6027\u5730\u64f4\u589e\u3002 \u53ef\u52a0\u5165\u6c2f\u9ef4\u7d20(Chloramphenicol)\u4ee5\u6291\u5236\u5bbf\u4e3b\u5408\u6210(host synthesis)\u4e26\u9632\u6b62\u7d30\u83cc\u67d3\u8272\u9ad4(bacterial chromosome)\u7684\u8907\u88fd\u3002LB Agar\uff0cMiller\u542b\u670910 g \/ L\u7684\u6c2f\u5316\u9209\uff0c\u8207Lennox\u914d\u65b9\u4e2d\u7684\u542b\u91cf\u4e0d\u540c\u3002 \u9019\u5141\u8a31\u7814\u7a76\u4eba\u54e1\u70ba\u7279\u5b9a\u83cc\u682a\u9078\u64c7\u6700\u4f73\u9e7d\u6fc3\u5ea6\u3002 \u57f9\u990a\u57fa\u53ef\u4ee5\u7121\u83cc\u88dc\u5145\u8461\u8404\u7cd6glucose\u6216\u6297\u751f\u7d20antibiotics\u3002\u8a3b\u660e: Product was previously known as Acumedia Product: 7213 LB Agar (Miller); and LabM Product: LAB168 LB Agar (Miller)\u3002\u6c2f\u5316\u9209\u7684\u91cf\u7d93\u5e38\u96a8\u7d30\u83cc\u7684\u751f\u9577\u9700\u6c42\u4e0d\u540c\u800c\u6709\u6240\u6539\u8b8aLB-Miller (10 g\/L NaCl)\u3001LB-Lennox (5 g\/L NaCl)\u3001LB-Luria (0.5 g\/L NaCl)<\/p>\n

\u80f0\u86cb\u767d\u8156 (Tryptone )————————————- 10.0 g \/ L<\/p>\n

\u9175\u6bcd\u62bd\u63d0\u7269(Yeast Extract ) ———-5.0g \/ L<\/p>\n

Sodium Chloride ————————10.0g \/ L<\/p>\n

Agar _________________15.0 g\/L<\/p>\n","protected":false},"excerpt":{"rendered":"

ProClone\u2122 \u9ad8\u6548\u80fdCompetent cell\u52dd\u4efb\u7d30\u80de | ABMGOOD\u8ca8\u865fE003 \u5177\u5099\u6709\u9ad8\u6548\u80fd\u7684 […]<\/p>\n","protected":false},"author":3,"featured_media":28511,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[5373],"tags":[6080],"_links":{"self":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/44527"}],"collection":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=44527"}],"version-history":[{"count":6,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/44527\/revisions"}],"predecessor-version":[{"id":46739,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/44527\/revisions\/46739"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/28511"}],"wp:attachment":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=44527"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=44527"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=44527"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}