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onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div><div id="liuh2dw" class="smallimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253047636719/652001.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div></div><div id="fewdntl" class="content"><p><span>需求數(shù)量：</span>0</p><p><span>價格要求：</span>面議</p><p><span>所在地：</span>上海市</p><p><span>包裝要求：</span><span style="color: #f30;"></span></p><p><span>產(chǎn)品關(guān)鍵詞：</span>上海食品安全熒光定量PCR實驗技術(shù)服務(wù),熒光定量PCR</p><p><span>***更新：</span>2020-08-14 05:05:56</p><p><span id="click" >瀏覽次數(shù)：0次</span></p><a rel="nofollow" class="button">聯(lián)系我們</a></div></div><div id="bac87mo" class="rightbox"><h3>公司基本資料信息</h3><p style="border:0;"><a rel="nofollow">上海朝瑞生物科技有限公司</a></p><p><span>聯(lián)系人：</span>仲先生</p><p><span>郵箱: </span>scienceladder@163.com</p><p><span>電話: </span>18621867065</p><p><span>傳真: </span>021_</p><p><span>網(wǎng)址: </span></p><p><span>手機: </span>021-54133071</p><p><span>地址: </span>上海市松江區(qū)廣富林路697號昂立大廈2113室</p><p>[當前離線] <a rel="nofollow"> [加為商友] </a><a rel="nofollow"> [發(fā)送信件] </a></p></div></div><div id="8npwiz3" class="page-proshow02"><p class="title">詳細說明</p><div id="nhtv3gn" class="box"><p><p style="text-indent:25px">TaqMan探針法：TaqMan 探針法是具有高度特異性的定量 PCR 技術(shù)。它的工作原理是在 PCR 反應(yīng)體系中存在一對 PCR 引物和一條探針，探針的5′ 端標記有報告基團，3′ 端標記有熒光淬滅基團，探針只與模板特異結(jié)合，其結(jié)合位點在兩條引物之間，上海食品安全熒光定量PCR實驗技術(shù)服務(wù)。當探針完整的時候，報告基團的熒光能量被淬滅基團吸收，上海食品安全熒光定量PCR實驗技術(shù)服務(wù)，上海食品安全熒光定量PCR實驗技術(shù)服務(wù)，所以儀器搜集不到信號，隨著反應(yīng)的進展，Taq酶遇到探針，利用 3′→5′ 外切核酸酶的活性把探針切斷，導致報告基團的熒光能量不能被淬滅基團吸收，產(chǎn)生了熒光信號，因此信號的強度就**了模板 DNA 的拷貝數(shù)上海食品安全熒光定量PCR實驗技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海食品安全熒光定量PCR實驗技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253047636719/652001.png"></div><p style="text-indent:25px">一般而言，熒光擴增曲線可以分成三個階段：熒光背景信號階段、熒光信號**擴增階段和平臺期。在熒光背景信號階段，擴增的熒光信號被熒光背景信號所掩蓋，無法判斷產(chǎn)物量的變化;在平臺期，擴增產(chǎn)物已不再呈**級增加，PCR終產(chǎn)物量與起始模板量之間沒有線性關(guān)系，無法計算起始模板拷貝數(shù)。因此只有在熒光信號**擴增階段，PCR產(chǎn)物量的對數(shù)值與起始模板量之間存在線性關(guān)系，故選擇在這個階段進行定量分析。為了定量方便，在實時熒光定量 PCR 技術(shù)中引入了幾個重要的概念：擴增曲線、熒光閾值、CT值。上海食品安全熒光定量PCR實驗技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海食品安全熒光定量PCR實驗技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046357422/4616552.png"></div><p> 熒光信號**擴增階段：只有在熒光產(chǎn)生進入**期， PCR產(chǎn)物量的對數(shù)值與起始模板量之間存在線性關(guān)系，所以在PCR反應(yīng)處于**期的某一點上來檢測PCR產(chǎn)物的量，由此來推斷模板**初的含量而進行定量分析。研究表明，每個模板的Ct值與該模板的起始拷貝數(shù)的對數(shù)存在線性關(guān)系，起始拷貝數(shù)越多，Ct值越小。通過已知起始拷貝數(shù)的標準品可得到標準曲線，只要獲得未知樣品的Ct值，即可從標準曲線上計算出該樣品的起始拷貝數(shù) </p> <p> 在平臺期，擴增產(chǎn)物已不再呈**級的增加，所以反應(yīng)終產(chǎn)物量與起始模板量之間已經(jīng)不存在線性關(guān)系，通過反應(yīng)終產(chǎn)物也算不出起始DNA拷貝數(shù) </p><p> 熒光定量PCR實驗步驟： </p> <p> 取凍存已裂解的細胞，室溫放置5分鐘使其完全溶解。 </p> <p> 兩相分離每1ml的TRIZOL試劑裂解的樣品中加入0.2ml的lvfang，蓋緊管蓋。手動劇烈振蕩管體15秒后，15到30℃孵育2到3分鐘。4℃下12000rpm離心15分鐘。離心后混合液體將分為下層的紅色酚lvfang相，中間層以及無色水相上層。RNA全部被分配于水相中。水相上層的體積大約是勻漿時加入的TRIZOL試劑的60%。 </p> <p> RNA沉淀將水相上層轉(zhuǎn)移到一干凈無RNA酶的離心管中。加等體積異丙醇混合以沉淀其中的RNA，混勻后15到30℃孵育10分鐘后，于4℃下12000rpm 離心10分鐘。此時離心前不可見的RNA沉淀將在管底部和側(cè)壁上形成膠狀沉淀塊。 </p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海食品安全熒光定量PCR實驗技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046611328/9318580.png"></div><p style="text-indent:25px">有關(guān)轉(zhuǎn)基因及其產(chǎn)品的安全性的爭論在國內(nèi)外愈演愈烈，各國**紛紛出臺一系列的政策、法規(guī)，要求對轉(zhuǎn)基因作物進行標識，有些國家對轉(zhuǎn)基因的比較低含量做了限定，其閾值大小從1-5％不等。這就要求必須對轉(zhuǎn)基因及其產(chǎn)品進行檢測。常規(guī)的檢測方法多采用PCR法，如PCR-凝膠電泳、PCR-ELISA等。這些方法普遍存在著PCR產(chǎn)物污染、特異性不高及速度慢等問題。熒光PCR技術(shù)是一種新近發(fā)展起來的檢測技術(shù)，它的應(yīng)用將從根本上解決這些問題。 1.收集了國內(nèi)外商品化的轉(zhuǎn)基因作物品種，并通過查閱有關(guān)文獻及網(wǎng)站初步確定了各種作物的轉(zhuǎn)基因構(gòu)建圖； 2．于反應(yīng)體系中引入UNG去污染酶，建立了無污染熒光PCR擴增體系； 3．以植物內(nèi)源基因18SrRNA為靶標，設(shè)計了特異性引物和熒光探針，建立了以18SrRNA基因的擴增與否來判斷核酸提取質(zhì)量好壞的方法； 4．以FAM熒光素為標記，建立了以35S、NOS、NPTⅡ、Pat、Bar、FMV和CryⅢa為篩選目標的轉(zhuǎn)基因農(nóng)作物熒光PCR定性檢測體系； 5．分別以FAM和TET兩種熒光素標記外源基因和內(nèi)源基因，建立了轉(zhuǎn)基因大豆及轉(zhuǎn)基因玉米的熒光定量PCR檢測體系。上海食品安全熒光定量PCR實驗技術(shù)服務(wù)</p><p style="text-indent:25px">上海食品安全熒光定量PCR實驗技術(shù)服務(wù)</p><p style="text-indent:25px">使用可高壓處理的移液器，由于移液器容易受產(chǎn)物氣溶膠或標本 DNA 的污染，因此要使用可高壓處理的移液器。嚴格按照試驗的分區(qū)進行操作，按照提取區(qū)、擴增區(qū)、檢測區(qū)單方向流動，物品、樣品和試劑不可逆向流動。操作多份樣品時，制備反應(yīng)混合液，先將熒光 PCR 反應(yīng)液、熒光探針和酶混合好，然后分裝到每個 PCR 管中，這樣即可以減少操作次數(shù)，避免污染，又可以增加反應(yīng)的精確度。加樣品、加蛋白酶 K 和加樣品過程中，特別注意防止樣品的交叉污染和酶的降解。操作時設(shè)立陰性及陽性對照，可驗證熒光 PCR反應(yīng)的準確性和可信性上海食品安全熒光定量PCR實驗技術(shù)服務(wù)</p><p style="text-indent:25px">上海朝瑞生物科技有限公司位于上海市，創(chuàng)立于2003-01-22。公司業(yè)務(wù)分為[ "科研技術(shù)服務(wù)", "應(yīng)用技術(shù)服務(wù)", "科研成果轉(zhuǎn)化", "科學產(chǎn)品銷售" ]等，目前不斷進行創(chuàng)新和服務(wù)改進，為客戶提供質(zhì)量的產(chǎn)品和服務(wù)。公司將不斷增強企業(yè)核心競爭力，努力學習行業(yè)先進知識，遵守行業(yè)規(guī)范，植根于化工行業(yè)的發(fā)展。公司自成立以來發(fā)展迅速，業(yè)務(wù)不斷發(fā)展壯大，年營業(yè)額度達到2000-3000萬元，未來我們將不斷進行創(chuàng)新和改進，讓企業(yè)發(fā)展再上新高。</p></p><br /><p>文章來源地址: http://www.cdcfah.com/cp/909583.html</p></div></div></div></div></div><div id="sxuwypm" class="page-right twoColRt"><div id="wb72krt" class="box01 margin10 clearfix"><h3><a href="http://www.cdcfah.com/cp/">猜你喜歡</a></h3><div id="laxzw2l" class="recomPic likePro"><a href="http://www.cdcfah.com/cp/814588.html"><div><img src="http://tyunfile.71360.com/UpLoadFile/2019/11/1/14/637082168864691056_lgpj_5374157.jpg" alt="福建ZF配件哪家強信息推薦自業(yè)物資供應(yīng)" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></div><p>福建ZF配件哪家強信息推薦自業(yè)物資供應(yīng)</p></a></div><div id="qf2fx8d" class="recomPic likePro"><a href="http://www.cdcfah.com/cp/846569.html"><div><img 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