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id="hyk2axe" class="twoCol"><div id="f2lxpb2" class="page-proshow clearfix"><p class="current"> 當(dāng)前位置:<a href="http://www.cdcfah.com/">首頁?<a href="http://www.cdcfah.com/cp/">產(chǎn)品供應(yīng)</a>?<a href="http://www.cdcfah.com/cp/second-_nongye/">農(nóng)業(yè)</a>?<a href="http://www.cdcfah.com/cp/third-nongye_nlmyxmhz/">農(nóng)林牧漁項(xiàng)目合作</a>?北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù) 歡迎咨詢上海朝瑞生物科技供應(yīng)</p><div id="l9xzgd7" class="leftbox clearfix"><h1>北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù) 歡迎咨詢上海朝瑞生物科技供應(yīng)</h1><div id="tplvwik" class="proBig"><div id="dzgik8b" class="bigimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253049365234/1538935.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div><div id="dsu78vs" class="smallimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253049365234/1538935.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div></div><div id="rqspbfb" class="content"><p><span>需求數(shù)量：</span>0</p><p><span>價(jià)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>北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù),熒光定量PCR</p><p><span>***更新：</span>2020-06-15 14:20:54</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="3lpwy8g" 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(jī): </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(dāng)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="ti2nkev" class="page-proshow02"><p class="title">詳細(xì)說明</p><div id="7yfcels" class="box"><p><p> 管家基因反應(yīng)體系：序號反應(yīng)物劑量 1 SYBR Green 1 染料 10μl 2 內(nèi)參照上游引物F 0.5μl 3 內(nèi)參照下游引物R 0.5μl 4 dNTP 0.5μl 5 Taq酶 1μl 6 待測樣品cDNA 5μl 7 ddH2O 32.5μl 8 總體積 50μl 輕彈管底將溶液混合，6000rpm短暫離心。 </p> <p> 制備好的陽性標(biāo)準(zhǔn)品和檢測樣本同時(shí)上機(jī)，反應(yīng)條件為：93℃2分鐘，然后93℃ 1分鐘，55℃ 2分鐘，共40個(gè)循環(huán)。針對每一需要測量的基因，選擇一確定表達(dá)該基因的cDNA模板進(jìn)行PCR反應(yīng)，北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)。 </p> <p> 反應(yīng)體系：序號反應(yīng)物劑量 1 10×PCR緩沖液 2.5 ul 2 MgCl2溶液 1.5 ul 3 上游引物F 0.5 ul 4 下游引物R 0.5 ul 5 dNTP混合液 3 ul 6 Taq聚合酶 1 ul 7 cDNA 1 ul 8 加水至總體積為 25ul 輕彈管底將溶液混合，6000rpm短暫離心。 </p> <p> 35個(gè)PCR循環(huán)(94℃1分鐘;55℃1分鐘;72℃1分鐘); 72℃延伸5分鐘，北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)。 </p> <p> PCR產(chǎn)物與 DNA Ladder在2%瓊脂糖凝膠電泳，溴化乙錠染色，檢測PCR產(chǎn)物是否為單一特異性擴(kuò)增條帶。 </p> <p> 將PCR產(chǎn)物進(jìn)行10倍梯度稀釋: 設(shè)定PCR產(chǎn)物濃度為1×1010，依次稀釋至109、108、107，北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)、106、105、104幾個(gè)濃度梯度。所有cDNA樣品分別配置實(shí)時(shí)定量 PCR反應(yīng)體系。北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253049365234/1538935.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(jìn)行標(biāo)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(jìn)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(xiàn)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U(xiǎn)NG去污染酶，建立了無污染熒光PCR擴(kuò)增體系； 3．以植物內(nèi)源基因18SrRNA為靶標(biāo)，設(shè)計(jì)了特異性引物和熒光探針，建立了以18SrRNA基因的擴(kuò)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(biāo)記，建立了以35S、NOS、NPTⅡ、Pat、Bar、FMV和CryⅢa為篩選目標(biāo)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(biāo)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。北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253043593750/8100830.png"></div><p> 實(shí)時(shí)熒光定量常用Taqman探針法和SYBR Green I法兩類。 </p> <p> 1.TaqMan探針法 </p> <p> TaqMan 探針法是具有高度特異性的定量PCR技術(shù)。它的工作原理為有一對 PCR 引物和一條探針存在于PCR 反應(yīng)體系中，有熒光淬滅基團(tuán)存在于3′ </p> <p> 端標(biāo)記，有報(bào)告基團(tuán)存在于探針的5′ 端標(biāo)記，探針*和模板特異結(jié)合，兩條引物之間是其結(jié)合點(diǎn)。因此信號儀器搜集不到，伴隨反應(yīng)的進(jìn)展，探針在外切核酸酶的活性的作用下被切斷，從而造成淬滅基團(tuán)不能吸收基團(tuán)的熒光能量，從而使熒光信號產(chǎn)生，所以模板 DNA 的拷貝數(shù)取決于信號的強(qiáng)度。 </p> <p> 2.SYBR Green I法 </p> <p> SYBR Green I為一種具有綠色激發(fā)波長的染料，其發(fā)射波長比較大約為 520 nm，比較大吸收波長約為 497 nm，能夠結(jié)合所有的雙螺旋小溝區(qū)域。由于處于游離狀態(tài)，SYBR Green I發(fā)出的熒光較弱，然而其結(jié)合雙鏈 DNA之后，就會使得熒光**增強(qiáng)，熒光信號的增加完全同步PCR 產(chǎn)物的增加。 </p><p> 產(chǎn)生的 ΔRn 值也不同。請注意，熒光信號基線屬于非模板依賴性因素，兩種預(yù)混液的基線是不同的（圖 3A）。Ct 值的變化并沒有反映出反應(yīng)系統(tǒng)的整體性能（圖 3B）。靈敏度相當(dāng)?shù)念A(yù)混液產(chǎn)生的 Ct ***值可能不同。 </p> <p> 使用預(yù)混液 A 和預(yù)混液 B 在等量的人類 gDNA 中進(jìn)行 RNase P 擴(kuò)增。(A) Rn 根據(jù)循環(huán)數(shù)進(jìn)行繪制，并且顯示兩個(gè)反應(yīng)的基線。(B) 對數(shù) (ΔRn) 根據(jù)循環(huán)數(shù)進(jìn)行繪制。兩種預(yù)混液的閾值（綠線）設(shè)定為相同水平。對于相同濃度的靶標(biāo)而言，預(yù)混液 B 的 Ct 值 (CtB) 早于預(yù)混液 A 的相應(yīng)值 (CtA)，表明預(yù)混液 B 的基線較低。使用 Applied Biosystems? 7500 實(shí)時(shí)熒光定量 PCR 系統(tǒng)執(zhí)行所有擴(kuò)增。 </p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046611328/9318580.png"></div><p> 實(shí)時(shí)熒光定量PCR的應(yīng)用 </p> <p> 目前，qPCR技術(shù)已經(jīng)廣泛應(yīng)用于生物醫(yī)藥食品等行業(yè)，應(yīng)用于疾病的早期診斷、遺傳病的早期診斷、***研究、**的診斷與研究、食品病原微生物的檢測、轉(zhuǎn)基因食品檢測、動物疫病檢測等，除此之外，還包括： </p> <p> ● 基因擴(kuò)增 </p> <p> ● 擴(kuò)增特異性分析 </p> <p> ● 基因定量分析 </p> <p> ● 基因檢測 </p> <p> ● 基因分型 </p> <p> ● SNP分析 </p> <p> ● RFLP多態(tài)性分析 </p> <p> ● 單/多基因表達(dá)研究 </p> <p> ● 高通量基因表達(dá)譜研究 </p> <p> 實(shí)時(shí)熒光定量PCR技術(shù)，顧名思義，就是在PCR的基礎(chǔ)上加入熒光基團(tuán)，通過熒光信號的變化的實(shí)時(shí)監(jiān)測PCR的反應(yīng)過程，***通過對未知模板進(jìn)行定量分析的方法。北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)</p><p style="text-indent:25px">北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)</p>熒光定量PCR(Flurogenic Quantitative Polymerase ChainReaction，F(xiàn)Q-PCR；real-time quantitative PCR, RT-qPCR or qPCR)，是1996 年由美國Applied Biosystems 公司推出的一種新定量試驗(yàn)技術(shù)，它是通過熒光染料或熒光標(biāo)記的特異性探針，標(biāo)記PCR產(chǎn)物進(jìn)行實(shí)時(shí)監(jiān)測反應(yīng)，利用與之相適應(yīng)的軟件對產(chǎn)物進(jìn)行分析，計(jì)算待測樣品模板的初始濃度 <p> 實(shí)時(shí)熒光定量PCR（Real-TimePCR）技術(shù)是在PCR技術(shù)的基礎(chǔ)上發(fā)展起來的核酸定量技術(shù)，是1996年美國Applied </p> <p> Biosystems公司推出，它是通過在PCR反應(yīng)體系中加入熒光基團(tuán)，然后利用熒光信號的積累強(qiáng)弱實(shí)時(shí)監(jiān)測整個(gè)PCR進(jìn)程，通過標(biāo)準(zhǔn)曲線對未知的DNA模板進(jìn)行定量。北京食品安全熒光定量PCR實(shí)驗(yàn)技術(shù)服務(wù)</p><p style="text-indent:25px">上海朝瑞生物科技有限公司致力于化工，以科技創(chuàng)新實(shí)現(xiàn)***管理的追求。公司自2003-01-22成立以來，投身于[ "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(xué)產(chǎn)品銷售" ]，是化工的主力軍。公司堅(jiān)持以技術(shù)創(chuàng)新為發(fā)展引擎，以客戶滿意為動力，目前擁有5~10人專業(yè)人員，年?duì)I業(yè)額達(dá)到2000-3000萬元。上海朝瑞科技始終關(guān)注化工市場，以敏銳的市場洞察力以準(zhǔn)確定位，實(shí)現(xiàn)與客戶的成長共贏。</p></p><br /><p>文章來源地址: http://www.cdcfah.com/cp/268024.html</p></div></div></div></div></div><div id="0holx7a" class="page-right twoColRt"><div id="hhtqsz8" class="box01 margin10 clearfix"><h3><a href="http://www.cdcfah.com/cp/">猜你喜歡</a></h3><div id="5zv2vnp" 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(qiáng) 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(qiáng) 信息推薦自業(yè)物資供應(yīng)</p></a></div><div id="08hoaho" 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