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基因組文庫(Genomic Library)構(gòu)建

時(shí)間:2015/7/1閱讀:504
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關(guān)鍵詞:基因組文庫(Genomic Library)構(gòu)建|實(shí)驗(yàn)技術(shù)服務(wù)
簡(jiǎn)介:世界*品牌基因組文庫(Genomic Library)構(gòu)建|實(shí)驗(yàn)技術(shù)服務(wù)原裝,,質(zhì)量保證,*。
基因組文庫(Genomic Library)構(gòu)建|實(shí)驗(yàn)技術(shù)服務(wù)——pCzn1質(zhì)粒+Arctic Express宿主菌低溫表達(dá)體系,。
我們具備豐富的蛋白表達(dá)設(shè)計(jì)經(jīng)驗(yàn)及實(shí)驗(yàn)操作技巧,承諾客戶不成功不收費(fèi)。我們所有的實(shí)驗(yàn)數(shù)據(jù)真實(shí)可信,,我們提供原始的表達(dá)菌株和克隆質(zhì)粒,,客戶可以依據(jù)我們的實(shí)驗(yàn)報(bào)告重復(fù)我們的實(shí)驗(yàn)結(jié)果。
產(chǎn)品品牌:基因組文庫(Genomic Library)構(gòu)建|實(shí)驗(yàn)技術(shù)服務(wù)   http://www.,。,。com/goodsid/fenleiyi/2868603/1.html    
測(cè)序?qū)嶒?yàn)流程:
基因組DNA文庫包括了基因組所有順序的隨機(jī)克隆片段。一個(gè)好的基因組DNA文庫的大小應(yīng)足夠大,,以便包括所有的基因DNA順序,。DNA克隆片段也應(yīng)足夠大,其中包括整個(gè)基因,、連接基因及它們穩(wěn)定形式所要的側(cè)翼順序,。為了獲得高的克隆效率和較大的插入片段,λ噬菌體載體和質(zhì)粒經(jīng)常被使用構(gòu)建基因組DNA文庫,。λ噬菌體文庫易于操作,,噬菌體載體上有多克隆位點(diǎn)。λ噬菌體能容納10-50kb插入DNA,,載體類型應(yīng)根據(jù)所要基因組DNA片的大小和用途來選擇,。
構(gòu)建一個(gè)基因組DNA文庫的基本步驟:
①分離純化基因組DNA;
②切割DNA成合適大小的片段以用于克??;
③載體DNA的制備;
④把基因組DNA片段和載DNA連接,;
⑤包裝重組分子,;
⑥測(cè)定入噬菌體滴度;
⑦擴(kuò)增DNA文庫,;
⑧評(píng)估文的質(zhì)量,。
基因組文庫構(gòu)建步驟:1、載體: 
l噬菌體:48.5kb,插入型(zui多可容納9kb),、取代型(可容納 38-51kb,,zui適19-20kb), EMBL, Charon 粘性質(zhì)粒(Cosmid):4-6kb, 質(zhì)粒+噬菌體的性質(zhì),,可容納大片段的外源基因,,zui多可容納46kb。 酵母人工染色體克隆體系(YAC,,Yeast Artificial Chromosome  Cloning System) 插入大小200—1000kb 2,、 載體DNA的制備: 高分子量的外源DNA 100-150kb載體DNA酶切反應(yīng)-----載體臂的純化-----載體脫磷處理 www.cqwestern。,。com(常用BamHI)   (蔗糖密度梯度離心)  (堿性磷酸酶) 3,、連接和包裝:外源片段與兩臂之間的比例,,包裝蛋白 4、感染宿主菌:選擇合適的宿主菌,,LE392, NM538, NM539, KW251等 5,、基因組文庫的保存和擴(kuò)增: 6、測(cè)滴度,,要在106fu以上
7,、保存:氯仿4℃,7% DMSO -70℃

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