{"id":6156,"date":"2016-08-31T14:21:04","date_gmt":"2016-08-31T06:21:04","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=6156"},"modified":"2022-07-26T14:43:22","modified_gmt":"2022-07-26T06:43:22","slug":"lentivirus%e6%85%a2%e7%97%85%e6%af%92rt-qpcr%e5%ae%9a%e9%87%8f%e4%b8%80%e5%80%8b%e6%ad%a5%e9%a9%9f%e5%bf%ab%e9%80%9frna%e8%90%83%e5%8f%96%e5%8f%8aqpcr","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=6156","title":{"rendered":"Lentivirus\u6162\u75c5\u6bd2RT-QPCR\u5b9a\u91cf|\u4e00\u500b\u6b65\u9a5f\u5feb\u901fRNA\u8403\u53d6\u53caQPCR"},"content":{"rendered":"

Lentivirus\u6162\u75c5\u6bd2\u7cfb\u5217\u5546\u54c1:<\/span> Lentivirus\u6162\u75c5\u6bd2\u4f4e\u901f30\u5206\u7d14\u5316\u5957\u7d44(\u7d14\u5316)<\/a> | Lentivirus \u6162\u75c5\u6bd2\u9ad8\u6fc3\u5ea6\u7d14\u5316\u5957\u7d44(\u7d14\u5316) <\/a>\u00a0| Lentivirus\u75c5\u6bd2\u5b9a\u91cf\u5feb\u7be9\u7247(\u5b9a\u91cf)<\/a> | Lentivirus\u75c5\u6bd2\u4e00\u500b\u6b65\u9a5fQPCR\u5b9a\u91cf\u5957\u7d44(\u5b9a\u91cf)<\/a><\/span><\/p>\n

\u3010<\/strong>\u75c5\u6bd2<\/strong>RT-QPCR<\/strong>\u5b9a\u91cf\u3011<\/strong>ABM qPCR Lentivirus Titration(Titer) Kit<\/strong>\u00a0Lentivirus<\/strong>\u75c5\u6bd2\u4e00\u500b\u6b65\u9a5fQPCR\u5b9a\u91cf\u5957\u7d44(\u5b9a\u91cf) (Cat. No. LV900)<\/strong><\/p>\n

Abmgood\u63d0\u4f9b\u5feb\u901f\u4e14\u65b9\u4fbf\u4e4bLentivirus\u75c5\u6bd2\u529b\u50f9\u5b9a\u91cf\u5957\u7d44\u3002\u5176\u7d50\u5408\u4e00\u500b\u6b65\u9a5f\u5feb\u901fRNA\u8403\u53d6\u53caQPCR\u6280\u8853 (SYBR system)\uff0c2\u500b\u5c0f\u6642\u5b8c\u6210\u5b9a\u91cf\u3002\u5957\u7d44\u5167\u9644\u6709reagent mix (reagent mix included with the kit includes primers and reverse transcriptase for RT) \u53caprimer\u3002primer\u4ee5HIV based vetors\u70ba\u8a2d\u8a08\u7684\u5075\u6e2c\u65b9\u5f0f (\u8a66\u5291\u4e0d\u5305\u62ecqPCR Mastermix, \u82e5\u9700\u9078\u64c7\u542bqPCR Mastermix\u4e4bLentivirus\u75c5\u6bd2\u529b\u50f9\u5b9a\u91cf\u5957\u7d44, \u8ca8\u865f\u70ba LV900-S\uff0cLV900-iC\uff0cLV900-R\u548cLV900-LR) \u3002\u5feb\u901f\u5b8c\u6210\u75c5\u6bd2\u529b\u50f9\u5b9a\u91cf\u5206\u6790\u3002\u53ef\u76f4\u63a5\u61c9\u7528\u65bc\u6e90\u65bc\u7522\u751f\u75c5\u6bd2\u7684\u7d30\u80de(virus-producing cell line)\u6216\u7d14\u5316\u7684\u75c5\u6bd2\u6a23\u54c1(purified viral preparations) \u4e4b\u75c5\u6bd2\u6548\u50f9(titer) \u78ba\u5b9a\u3002\u8a66\u5291\u76d2\u4e2d\u63d0\u4f9b\u75c5\u6bd2\u88c2\u89e3\u7de9\u885d\u6db2(Virus Lysis Buffer) \u6709\u52a9\u65bc\u63d0\u53d6\u4f86\u81ea\u75c5\u6bd2\u6a23\u54c1\u7684RNA\uff08extract viral RNA\u75c5\u6bd2\u88c2\u89e3\u6db2\uff09\u3002\u7136\u5f8c\u5c07Reagentmix\u548cLentivirus 2X qPCR MasterMix\u8207\u75c5\u6bd2\u88c2\u89e3\u6db2\u548c\u6162\u75c5\u6bd2\u6a19\u6e96\u54c1\uff08STD1 \/ STD2\uff09\uff0c\u53ef\u901a\u904eqRT-PCR\u78ba\u5b9a\u95be\u503c\u5faa\u74b0\uff08threshold cycle , Ct\uff09\u503c (comparative Ct experiment)\u3002\u7136\u5f8c\u5c07\u6240\u5f97\u7684Ct\u503c\u7528\u65bc\u8a08\u7b97\u75c5\u6bd2\u6a23\u54c1\u7684\u6162\u75c5\u6bd2\u529b\u50f9\u3002\u6b64\u5546\u54c1\u4e0d\u6703\u6709NTC\u91cf\u5316\u554f\u984c\uff0c\u5b8c\u5168\u6392\u9664NTC\u8a0a\u865f\u3002\u7cbe\u78ba\u5b9a\u91cfLentivirus titer\u4e4b\u65b9\u6cd5\u3002\u6a19\u6e96\u54c1STD\u70balentivirus\u75c5\u6bd2 (\u75c5\u6bd2\u8f09\u9ad4DNA)\u3002Standard\u6fc3\u5ea6\u70ba\u00a05x 10^7 IU\/ml. Standard 2\u00a0\u6fc3\u5ea6\u70ba6.25×10^6 IU\/ml.\u00a0\u5169\u8005standards\u00a0\u76f8\u5dee\u00a02^3,\u00a0\u76f8\u7576\u65bcCt\u00a0\u503c3\u6b21\u65b9\u3002\u6700\u4f4e\u6e2c\u91cf\"\"10^6<\/sup>IU\/ml \u3002Normally, for a viral sample containing around 10^6 IU\/ml, the Ct should be around the same as STD1 and STD2. If your Ct values for your samples are beyond 22, this indicates that the titre is too low to obtain accurate readings (6 Ct later than STD2 will indicate that your viral samples are 100X more diluted than Std2).<\/p>\n

\u60a8\u53ef\u4ee5\u4f7f\u7528\u6211\u5011\u7684\u6162\u75c5\u6bd2\u529b\u50f9\u8a08\u7b97\u5668\r\n<\/span><\/pre>\n
Average Ct of STD1\u00a0\u00a0\u00a0\u00a0\u00a0
\nAverage Ct of STD2\u00a0\u00a0\u00a0\u00a0\u00a0
\nAverage Ct of Sample\u00a0
\n\u00a0\u00a0<\/p>\n
<\/div>\n
Titer of sample:\u00a0<\/span>\u00a0IU\/ml<\/b><\/div>\n<\/form>\n
1) STD1, STD2 are provided in the Lentivirus qPCR Titer Kit.
\n2) The equation we use in this calculation of the titer tool contains a specific coefficient determined in-house to convert the value from GC\/ml to IU\/ml; as such, the final titer value obtained is in IU\/ml.<\/p>\n
    \n
  1. One-Step qPCR\uff0c2\u5c0f\u6642\u5167\u53ef\u5b8c\u6210\u5b9a\u91cf\u3002<\/li>\n
  2. Primer\u7d93\u7279\u6b8a\u8a2d\u8a08\uff0c\u89e3\u6c7aprimer dimer\u5e72\u64fe\u3002<\/li>\n
  3. \u9644\u6709\u75c5\u6bd2\u6fc3\u5ea6\u6a19\u6e96\u6db2\u5169\u7d44\uff0c\u6e96\u78ba\u5b9a\u91cf\u75c5\u6bd2\u6fc3\u5ea6\u3002<\/li>\n
  4. \u642d\u914dABM qPCR Mastermix<\/a><\/u>\uff0c\u6548\u679c\u66f4\u4f73\u3002( \u82e5\u9700\u9078\u64c7\u542bqPCR Mastermix\u4e4bLentivirus\u75c5\u6bd2\u529b\u50f9\u5b9a\u91cf\u5957\u7d44, \u8ca8\u865f\u70ba LV900-S\uff0cLV900-iC\uff0cLV900-R\u548cLV900-LR)TaqMan qPCR Mastermix\u7cfb\u5217\u4e0d\u9069\u7528\u65bcLV900-qPCR\u6162\u75c5\u6bd2\u6ef4\u5ea6\uff08\u6ef4\u5b9a\uff09\u8a66\u5291\u76d2\u3002\u8a72\u8a66\u5291\u76d2\u53ea\u80fd\u4f7f\u7528SybrGreen\u6216BrightGreen\u6210\u5206\u7684qPCR Mastermix\u3002<\/li>\n<\/ol>\n
    \u611f\u67d3\u8907\u6578\uff08MOI\uff09\u7684\u8a08\u7b97\u65b9\u6cd5\uff1aMOI =\uff08\u7522\u54c1\u6ef4\u5ea6\uff08IU \/ ml\uff09x\u75c5\u6bd2\u9ad4\u7a4d\uff08ml\uff09\uff09\/\u7e3d\u7d30\u80de\u6578\u4f8b\u5982\uff0c\u4ee51 x 10 ^ 6 IU \/ ml\u6548\u50f9\u7372\u5f9710\u7684MOI\u5982\u679c\u75c5\u6bd2\u57281 x 10 ^ 5\u500b\u7d30\u80de\u4e0a\uff0c\u8acb\u4f7f\u75281.0\u6beb\u5347\u75c5\u6bd2\u3002<\/p>\n\n\n\n\n\n\n\n
    \u7522\u54c1\u7de8\u865f<\/strong><\/td>\n\u7522\u54c1\u540d\u7a31<\/strong><\/td>\n\u5305\u88dd<\/strong><\/td>\n<\/tr>\n
    LV999<\/td>\nSpeedy Lentivirus Purification<\/a><\/td>\n100ml<\/td>\n<\/tr>\n
    LV900<\/td>\nqPCR Lentivirus Titration(Titer) Kit<\/a><\/td>\n100 Reactions<\/td>\n<\/tr>\n
    LV998<\/td>\nUltra-Pure Lentivirus Purification Kit<\/a>\uff0d\u53ef\u7d14\u5316\u75c5\u6bd2\u6fc3\u5ea6\u90541011<\/sup>\u75c5\u6bd2\u9846\u7c92<\/td>\n1 Kit<\/td>\n<\/tr>\n
    G171
    \nG172
    \nG173<\/td>\n    		PuRetro\u2122 Lentivirus Purification Kit<\/a><\/td>\n2 Standard Units
    \n5 Standard Units
    \n2 Giga Units<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
    \"\u672a\u547d\u540d-14\"\u3010<\/strong>\u75c5\u6bd2\u4f4e\u901f\u7d14\u5316\u3011<\/strong> ABM Speedy Lentivirus Purification kit <\/strong>\u6162<\/strong>\u75c5\u6bd2<\/strong>\u4f4e<\/strong>\u901f30<\/strong>\u5206<\/strong>\u7d14\u5316\u5957\u7d44 (100<\/strong>\u500d\u6fc3\u7e2e<\/strong>)<\/strong> \u00a0(Cat. No. LV999)<\/strong><\/p>\n
    \u50b3\u7d71\u7684\u75c5\u6bd2\u7d14\u5316\u65b9\u6cd5\uff0c\u5927\u591a\u5229\u7528\u8d85\u9ad8\u901f\u96e2\u5fc3(ultracentrifuge)\uff0c\u642d\u914d\u6fc3\u5ea6\u68af\u5ea6(CsCl)\u7b49\u65b9\u6cd5\u9032\u884c\u75c5\u6bd2\u5206\u96e2\uff0c\u9700\u8981\u7d14\u719f\u6280\u8853\u4e14\u904e\u7a0b\u8cbb\u6642\u3002Speedy Lentivirus purification kti\u75c5\u6bd2\u7d14\u5316\u5957\u7d44\u53ef\u514d\u9664\u4f7f\u7528\u8d85\u9ad8\u901f\u96e2\u5fc3\uff0c\u66f4\u7c21\u55ae\u3001\u5feb\u901f\u53d6\u5f97\u9ad8\u6fc3\u5ea6\u75c5\u6bd2\u6db2\u300230\u5206\u9418\u5167\u5feb\u901f\u7d14\u5316\u75c5\u6bd2\u3001\u53ef\u9054\u6fc3\u7e2e\u6548\u679c\u7d04100x\u3002<\/p>\n
    \u200b\"ABM_Speedy\"<\/p>\n
    293 cells \u611f\u67d3\u7d14\u5316\u524d\uff08A\uff09\u8207\u7d14\u5316\u5f8c\uff08B\uff09\u7684lentivirus 48hr\u5f8c\u7684\u7d50\u679c<\/p>\n
    \"\u672a\u547d\u540d-14\"\u3010\u75c5\u6bd2\u9ad8\u6fc3\u5ea6\u7d14\u5316 ++plus<\/sup>\u3011Ultra-Pure Lentivirus Purification Kit \u6162\u75c5\u6bd2\u7d14\u5316\u53ca\u6fc3\u5ea6\u5957\u7d44\uff0c\u6709\u6548\u63d0\u9ad8\u75c5\u6bd2\u7d14\u5ea6\u53ca\u529b\u50f9 (Cat. No. LV998)<\/strong><\/p>\n
    \u91cd\u7d44\u6162\u75c5\u6bd2lentivirus\uff0c\u5ee3\u6cdb\u61c9\u7528\u65bc\u7d30\u80de\u4e4b\u8f49\u5c0e\u3002\u8f49\u67d3\u5f8c\u7684\u75c5\u6bd2\u4e0a\u6e05\u6db2\u529b\u50f9\uff0c\u7d04106<\/sup> CFU\u83cc\u843d\u5f62\u6210\u55ae\u4f4d\uff08\u83cc\u843d\u5f62\u6210\u55ae\u4f4d\uff09\/ ml\u3002\u5728\u9ad8\u6548\u80fd\u75c5\u6bd2\u529b\u50f9\u53ca\u9ad4\u5167\u57fa\u56e0\u50b3\u905e\u4e2d\uff0c\u8981\u6c42\u8f03\u9ad8\u7684\u75c5\u6bd2\u7d14\u5ea6\u53ca\u529b\u50f9\uff0c\u4e14\u65bc\u6d3b\u9ad4\u8a66\u9a57\u4e2d\uff0c\u907f\u514d\u75c5\u6bd2\u7d30\u80de\u57f9\u990a\u4e0a\u6e05\u6db2\uff0c\u6240\u9020\u6210\u4e4b\u6c61\u67d3\u53ef\u80fd\u6027\u3002\u56e0\u6b64\uff0c\u75c5\u6bd2\u4e0a\u6e05\u9700\u8981\u9032\u4e00\u6b65\u6fc3\u7e2e\u53ca\u7d14\u5316\u3002Abmgood\u63d0\u4f9b\u4e00\u5957\u75c5\u6bd2\u7d14\u5316\u5957\u7d44\u3002\u50b3\u7d71\u7d14\u5316\u75c5\u6bd2\uff0c\u7d93\u7531\u8d85\u901f\u96e2\u5fc3\u6fc3\u7e2e\u3002\u96d6\u7136\u8a72\u65b9\u6cd5\u6548\u679c\u5f88\u597d\uff0c\u4f46\u9700\u8981\u7279\u5b9a\u7684\u7684\u8d85\u901f\u96e2\u5fc3\u8a2d\u5099\uff0c\u4e14\u6280\u8853\u8981\u6c42\u9ad8\u3002\u6b64\u5916\uff0c\u4ee5\u6fc3\u7e2e\u7684\u75c5\u6bd2\u4e0a\u6e05\u7e3d\u9ad4\u7a4d\u662f\u6709\u9650\u7684\u8d85\u901f\u96e2\u5fc3\u7ba1\u7684\u5927\u5c0f\u3002\u5831\u544a\u6307\u51fa\uff0c\u8d85\u901f\u96e2\u5fc3\u904e\u7a0b\u4e2d\uff0c\u6703\u9020\u6210\u4e00\u4e9b\u4e0d\u5229\u7684\u7269\u7406\u6548\u61c9\uff0c\u6703\u5f71\u97ff\u75c5\u6bd2\u9846\u7c92\u7684\u751f\u7269\u6d3b\u6027\u3002<\/p>\n
    Abmgood\u767c\u5c55 Ultra-Pure Lentivirus Purification Kit (Cat. No.LV998).<\/p>\n
    \u91dd\u5c0d\u75c5\u6bd2\u6fc3\u7e2e\u6280\u8853\u53ca\u7d14\u5ea6\u505a\u6539\u826f\u3002\u63d0\u4f9b\u4e00\u500b\u5feb\u901f\u4e14\u6709\u6548\uff0c\u7d14\u5316\u53ca\u6fc3\u7e2e\u75c5\u6bd2\u7684\u7522\u54c1\u3002\u5176\u8a2d\u8a08\u539f\u7406\uff0c\u70ba\u5229\u7528\u96e2\u5b50\u4ea4\u63db\u7684\u7d50\u5408\u548c\u7368\u7279\u7de9\u885d\u6db2\uff0c\u6700\u5927\u9650\u5ea6\u6e1b\u5c11\u75c5\u6bd2\u9846\u7c92\u7684\u640d\u58de\uff0c\u4e14\u63d0\u9ad8\u75c5\u6bd2\u529b\u50f9\u3002<\/p>\n
    \"\u672a\u547d\u540d-14\"\u3010\u75c5\u6bd2<\/strong>\u5feb\u7be9\u5b9a\u91cf<\/strong>\u3011<\/strong><\/strong>\u00a0Instant Lentivirus Titration Card<\/strong>\u6162\u75c5\u6bd2\u5b9a\u91cf\u5361\/<\/strong>\u5feb\u7be9\u5f0f\u6aa2\u6e2c<\/strong>(Cat. No. G614\/G615)<\/strong><\/p>\n
    \u00a0\"\"<\/strong><\/p>\n
    \u4f7f\u7528\u6a6b\u5411\u6d41\u65b9\u6cd5\uff08lateral flow immunoassay\uff09\u5075\u6e2clentiviral capsule protein\u9032\u884c\u5b9a\u91cf\u3002\u6bd4\u8d77\u50b3\u7d71infection based assays\u97003-4\u5929\uff0cInstant Lentivirus Titration Card \u53ea\u898110\u5206\u9418\u5373\u53ef\u5f97\u5230\u5b9a\u91cf\u7d50\u679c\u3002\u7c21\u55ae\u64cd\u4f5c\uff0c10\u5206\u9418\u7372\u5f97\u7d50\u679c\uff0d\u4e0d\u9700\u984d\u5916\u8403\u53d6\u6b65\u9a5f\uff1b\u76ee\u6e2c\u89c0\u5bdf\u7d50\u679c\u984f\u8272\u8b8a\u5316\u00a0\u7528\u9014\uff1a\u8a55\u4f30\u57f9\u990a\u6db2\u4e2d\u9069\u7576\u7684\u75c5\u6bd2\u6fc3\u5ea6\u3002\u9748\u654f\u5ea6\u53ef\u90541\u00d7105<\/sup>\u00a0IU\/mL\u75c5\u6bd2\u6fc3\u5ea6\uff08\u62167.7 pg\/ml\u7684p24 viral capsid protein\uff09<\/p>\n
    \"\u672a\u547d\u540d-14\"<\/strong><\/p>\n
    Lentivirus\u6162\u75c5\u6bd2\u4f4e\u901f30\u5206\u7d14\u5316\u5957\u7d44(\u7d14\u5316)<\/a> | Lentivirus \u6162\u75c5\u6bd2\u9ad8\u6fc3\u5ea6\u7d14\u5316\u5957\u7d44(\u7d14\u5316) <\/a>\u00a0| Lentivirus\u75c5\u6bd2\u5b9a\u91cf\u5feb\u7be9\u7247(\u5b9a\u91cf)<\/a> | Lentivirus\u75c5\u6bd2\u4e00\u500b\u6b65\u9a5fQPCR\u5b9a\u91cf\u5957\u7d44(\u5b9a\u91cf)<\/a><\/p>\n
     <\/p>\n","protected":false},"excerpt":{"rendered":"
    Lentivirus\u6162\u75c5\u6bd2\u7cfb\u5217\u5546\u54c1: Lentivirus\u6162\u75c5\u6bd2\u4f4e\u901f30\u5206\u7d14\u5316\u5957\u7d44(\u7d14\u5316) | Lentivi […]<\/p>\n","protected":false},"author":3,"featured_media":6157,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[57,456],"tags":[4568],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/6156"}],"collection":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=6156"}],"version-history":[{"count":3,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/6156\/revisions"}],"predecessor-version":[{"id":24365,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/6156\/revisions\/24365"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/6157"}],"wp:attachment":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=6156"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=6156"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=6156"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}