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Biosystems公司推出，由于該技術(shù)不僅實(shí)現了PCR從定性到定量的飛躍，而且與常規PCR相比，它具有特異性更強、有效解決PCR污染問(wèn)題、自動(dòng)化程度高等特點(diǎn)，目前已得到廣泛應用;實(shí)時(shí)熒光定量PCR：在PCR反應體系中加入熒光基團，利用熒光信號積累實(shí)時(shí)監測整個(gè)PCR進(jìn)程，Z后通過(guò)標準曲線(xiàn)對未知模板進(jìn)行定量分析的方法。</p></div> <ul> <li><img src="/Skins/270236/images/dot.png" /><span>產(chǎn)品型號：</span></li> <li><img src="/Skins/270236/images/dot.png" /><span>廠(chǎng)商性質(zhì)：</span>生產(chǎn)廠(chǎng)家</li> <li><img src="/Skins/270236/images/dot.png" /><span>更新時(shí)間：</span>2024-05-13</li> <li><img src="/Skins/270236/images/dot.png" /><span>訪(fǎng) 問(wèn) 量：</span>5088</li> </ul> <div id="im8tiuhso" class="pro_detail_btn"> <a href="#order" class="prodtl_btn1">產(chǎn)品咨詢(xún)</a><a href="/contact.html" target="_blank" class="prodtl_btn2">聯(lián)系我們</a> </div> </div> <div id="im8tiuhso" class="clear"></div> </div>  <div class="im8tiuhso" id="ny_pro_box02"> <div class="im8tiuhso" id="con"> <ul id="tags"> <li id="im8tiuhso" class=selectTag><A onmouseover="selectTag('tagContent0',this)" href="javascript:void(0)"onFocus="this.blur()">產(chǎn)品介紹：</A> </li> </ul> <div class="im8tiuhso" id=tagContent> <div id="im8tiuhso" class="tagContent selectTag" id=tagContent0><p><strong>實(shí)時(shí)熒光定量PCR技術(shù)（Real Time Quantitative PCR）</strong></p><p>在PCR反應體系中加入熒光基團，利用熒光信號積累實(shí)時(shí)監測整個(gè)PCR進(jìn)程，zui后通過(guò)標準曲線(xiàn)對未知模板進(jìn)行定量分析的方法。</p><p>相對定量應用（Relative Quantification，RQ） </p><p> 特異性熒光標記：</p><p> TaqMan法</p><p> TaqMan探針的特性：</p><p>（1）5′端標記有報告基團（Reporter, R），如FAM、VIC等</p><p>（2）3′端標記有熒光淬滅基團（Quencher, Q）</p><p>（3）探針完整，R所發(fā)射的熒光能量被Q基團吸收，無(wú)熒光， R與Q分開(kāi)，發(fā)熒光</p><p>（4）Taq酶有 5′→3′外切核酸酶活性，可水解探針</p><p><strong>相對定量通過(guò)內標定量：</strong><br />內標（Endogenous Control）通常是β-actin、GAPDH基因等看家基因，在細胞中的表達量或在基因組中的拷貝數恒定，受環(huán)境因素影響小，內標定量結果代表了樣本中所含細胞或基因組數量。</p><p><strong>相對定量分析方法：</strong></p><p> 2-ΔΔCt<br />前提：目標序列和內參序列的擴增效率接近100％且偏差在5％以?xún)?br /> 1、用內參基因的CT值歸一目標基因的CT值：<br /> ΔCT（test）= CT（target,test）-CT（ref,test） <br /> ΔCT（calibrator）= CT（target,calibrator）-CT（ref, calibrator） <br /> 2、用校準樣本的ΔCT值歸一試驗樣本的ΔCT值：<br /> ΔΔCT= ΔCT（test） - ΔCT（calibrator）<br /> 3、計算表達水平比率：<br /> 2 –ΔΔCT=表達量的比值</p><p><strong>服務(wù)流程：</strong></p><table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td><p>步驟</p></td><td><p>客戶(hù) 提供</p></td><td><p>服務(wù) 內容</p></td><td><p>基屹提供</p></td></tr><tr><td><p>1</p></td><td><p>新鮮的且盡量多的材料；已知的全長(cháng)基因序列；盡可能詳細的背景資料：DNA/ RNA來(lái)源、豐度等。</p></td><td><p>DNA/RNA提取，根據基因序列設計特異性的引物和熒光探針。</p></td><td> </td></tr><tr><td><p>2</p></td><td><p>純化好的DNA/RNA（大于5 ug/樣品）；已知的全長(cháng)基因序列；盡可能詳細的背景資料：DNA/ RNA來(lái)源、豐度等。</p></td><td><p>根據基因序列設計特異性的引物和熒光探針。</p></td><td> </td></tr><tr><td><p>3</p></td><td> </td><td><p>以DNA為模板，對目的基因進(jìn)行熒光定量PCR檢測分析</p></td><td><p>提供完整的定量分析結果、實(shí)驗熒光定量擴增曲線(xiàn)及詳細的實(shí)驗步驟。</p></td></tr></tbody></table><p>注意：客戶(hù)可根據所能提供的起始材料不同，選擇從步驟1或2啟動(dòng)項目。</p><p> </p><p> </p></div> <div class="im8tiuhso" id="nr_textbox"><link rel="stylesheet" type="text/css" 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