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class="smallimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046357422/4616552.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div></div><div id="geqxu3w" 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>江蘇轉(zhuǎn)基因熒光定量PCR課題承包,熒光定量PCR</p><p><span>***更新：</span>2020-08-06 20:13:52</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="hvsjqmd" 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(hào)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="r2wgnp2" class="page-proshow02"><p class="title">詳細(xì)說明</p><div id="yxeb3z3" class="box"><p><p style="text-indent:25px">SNP基因分型技術(shù)原理是PCR擴(kuò)增含有SNP的基因組片段，主要特點(diǎn)是準(zhǔn)確性高、靈活性強(qiáng)、通量大，主要方法是TaqMan探針法。TaqMan探針法針對(duì)染色體上的不同SNP位點(diǎn)分別設(shè)計(jì)PCR引物和TaqMan探針，進(jìn)行實(shí)時(shí)熒光PCR擴(kuò)增。探針的5’-端和3’-端分別標(biāo)記一個(gè)報(bào)告熒光基團(tuán)和一個(gè)淬滅熒光基團(tuán)。當(dāng)溶液中存在PCR產(chǎn)物時(shí)，該探針與模板退火，即產(chǎn)生了適合于核酸外切酶活性的底物，江蘇轉(zhuǎn)基因熒光定量PCR課題承包，江蘇轉(zhuǎn)基因熒光定量PCR課題承包，從而將探針5’-端連接的熒光分子從探針上切割下來，破壞兩熒光分子間的PRET，江蘇轉(zhuǎn)基因熒光定量PCR課題承包，發(fā)出熒光。通常用于少量SNP位點(diǎn)分析。江蘇轉(zhuǎn)基因熒光定量PCR課題承包</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇轉(zhuǎn)基因熒光定量PCR課題承包,熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046357422/4616552.png"></div><p style="text-indent:25px">實(shí)時(shí)熒光定量聚合酶鏈?zhǔn)椒磻?yīng)（PCR）指的是在PCR進(jìn)行的同時(shí)，對(duì)其過程進(jìn)行監(jiān)測(cè)的能力（即實(shí)時(shí)）。因此，數(shù)據(jù)可在PCR擴(kuò)增過程中，而非PCR結(jié)束之后，進(jìn)行收集。這為基于PCR的DNA和RNA定量方法帶來了**性的變革。在實(shí)時(shí)熒光定量PCR (qPCR)中，反應(yīng)以循環(huán)中***檢測(cè)到目標(biāo)擴(kuò)增的時(shí)間點(diǎn)為特征，而非在一定循環(huán)數(shù)后目標(biāo)分子累計(jì)的擴(kuò)增量。目標(biāo)核酸的起始拷貝數(shù)越高，則會(huì)越快觀察到熒光的***增加。相反，終點(diǎn)法檢測(cè)（也稱 “讀板檢測(cè)”）測(cè)量的是PCR循環(huán)結(jié)束時(shí)累計(jì)的PCR產(chǎn)物量。上海siRNA熒光定量PCR檢測(cè)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇轉(zhuǎn)基因熒光定量PCR課題承包,熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253043593750/2901163.png"></div><p> 在使用具有相同 ROX 水平的兩種預(yù)混液的一次檢測(cè)中獲得的原始熒光數(shù)據(jù)。信號(hào)差異是由預(yù)混液的成分造成的。使用 Applied Biosystems? VIC?/MGB 探針在 Applied Biosystems? 7900HT 快速實(shí)時(shí)熒光定量 PCR 系統(tǒng)上執(zhí)行此反應(yīng)。x 軸顯示熒光基團(tuán)的的發(fā)射波長(zhǎng)，y 軸顯示發(fā)射強(qiáng)度。 </p> <p> 任何分子的熒光發(fā)射都受環(huán)境因素的影響，比如溶液的 pH 值和鹽濃度。圖 2 顯示某個(gè) Applied Biosystems? TaqMan® 探針在兩種不同預(yù)混液背景下的原始熒光數(shù)據(jù)。請(qǐng)注意，盡管兩組反應(yīng)的靶標(biāo)、探針和 ROX 染料濃度都相同，預(yù)混液 A 中的熒光強(qiáng)度更高 </p><p style="text-indent:25px">有關(guān)轉(zhuǎn)基因及其產(chǎn)品的安全性的爭(zhēng)論在國(guó)內(nèi)外愈演愈烈，各國(guó)**紛紛出臺(tái)一系列的政策、法規(guī)，要求對(duì)轉(zhuǎn)基因作物進(jìn)行標(biāo)識(shí)，有些國(guó)家對(duì)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(duì)轉(zhuǎn)基因及其產(chǎn)品進(jìn)行檢測(cè)。常規(guī)的檢測(cè)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(cè)技術(shù)，它的應(yīng)用將從根本上解決這些問題。 1.收集了國(guó)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(cè)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(cè)體系。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇轉(zhuǎn)基因熒光定量PCR課題承包,熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253043593750/2359026.png"></div><p> 每個(gè)反應(yīng)管內(nèi)的熒光信號(hào)到達(dá)設(shè)定閾值時(shí)所經(jīng)歷的循環(huán)數(shù)。每個(gè)模板的Ct值與該模板的起始拷貝數(shù)的對(duì)數(shù)存在線性關(guān)系，起始拷貝數(shù)越多，Ct值越小。利用已知起始拷貝數(shù)的標(biāo)準(zhǔn)品可作出標(biāo)準(zhǔn)曲線，其中橫坐標(biāo)**起始拷貝數(shù)的對(duì)數(shù)，縱坐標(biāo)**Ct值。因此，只要獲得未知樣品的Ct值，即可從標(biāo)準(zhǔn)曲線上計(jì)算出該樣品的起始拷貝數(shù)。同時(shí)，對(duì)于熒光PCR實(shí)驗(yàn)來說，CT值也是判讀樣品陰陽性的重要指標(biāo)。 </p> <p> 大量的熒光定量PCR研究資料表明，病原體數(shù)量與***性疾病病情的輕重程度、傳染性及***效果均有相關(guān)性。因此，通過熒光定量PCR方法得到的檢測(cè)結(jié)果，一定程度上可以準(zhǔn)確反映疾病***的情況。江蘇lncRNA熒光定量PCR檢測(cè)服務(wù)</p><p style="text-indent:25px">江蘇轉(zhuǎn)基因熒光定量PCR課題承包</p><p> 熒光定量PCR 實(shí)驗(yàn)步驟（1） </p> <p> 樣品RNA的抽提 </p> <p> <br /> </p> <p> ①取凍存已裂解的細(xì)胞，室溫放置5分鐘使其完全溶解。 </p> <p> <br /> </p> <p> ②每樣品加入1ml的TRIZOL試劑裂解，蓋緊管蓋。手動(dòng)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(shí)加入的TRIZOL試劑的60%。 </p> <p> <br /> </p> <p> ③RNA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(shí)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> <p> <br /> </p> <p> ④RNA清洗移去上清液，每1mlTRIZOL試劑裂解的樣品中加入至少1ml的75%乙醇(75%乙醇用DEPCH2O配制)，清洗RNA沉淀?；靹蚝?，4℃下7000rpm離心5分鐘。 </p> <p> <br /> </p> <p> ⑤RNA干燥小心吸去大部分乙醇溶液，使RNA沉淀在室溫空氣中干燥5-10分鐘。 </p> <p> <br /> </p> <p> ⑥溶解RNA沉淀溶解RNA時(shí)，先加入無RNA酶的水40μl用移液器反復(fù)吹打幾次，使其完全溶解，獲得的RNA溶液保存于-80℃待用。江蘇轉(zhuǎn)基因熒光定量PCR課題承包</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)涵蓋[ "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" ]等，價(jià)格合理，品質(zhì)有保證。公司注重以質(zhì)量為中心，以服務(wù)為理念，秉持誠(chéng)信為本的理念，打造化工質(zhì)量品牌。公司憑借深厚技術(shù)支持，年?duì)I業(yè)額度達(dá)到2000-3000萬元，并與多家行業(yè)**公司建立了緊密的合作關(guān)系。</p></p><br /><p>文章來源地址: 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