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/><span>訪問量：</span>2417</li> </ul> <div id="x4vfcpe" class="pro_detail_btn"> <a href="#order" class="prodtl_btn1">產(chǎn)品咨詢</a><a href="/contact.html" target="_blank" class="prodtl_btn2">聯(lián)系我們</a> </div> </div> <div id="x4vfcpe" class="clear"></div> </div>  <div class="x4vfcpe" id="ny_pro_box02"> <div class="x4vfcpe" id="con"> <ul id="tags"> <li id="x4vfcpe" class=selectTag><A onmouseover="selectTag('tagContent0',this)" href="javascript:void(0)"onFocus="this.blur()">產(chǎn)品介紹：</A> </li> </ul> <div class="x4vfcpe" id=tagContent> <div id="x4vfcpe" class="tagContent selectTag" id=tagContent0><strong> <span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">熒光定量PCR技術(shù)</span></span></span></strong><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">與普通PCR的區(qū)別：</span></span></span><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　理論上PCR是一個指數(shù)增長的過程，但是實際的PCR擴增曲線并不是標準的指數(shù)曲線，而是S形曲線。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　這是因為隨著PCR循環(huán)的增多，擴增規(guī)模迅速增大，Taq酶、dNTP、引物，甚至DNA模板等各種PCR要素逐漸不敷需求，PCR的效率越來越低，產(chǎn)物增長的速度就逐漸減緩。當所有的Taq酶都被飽和以后，PCR就進入了平臺期。由于各種環(huán)境因素的復(fù)雜相互作用，不同的PCR反應(yīng)體系進入平臺期的時機和平臺期的高低都有很大變化，難以精確控制。所以，即使是96次PCR重復(fù)實驗，各種條件基本*，所得結(jié)果有很大差異，所以在實驗過程中我們不能根據(jù)zui終PCR產(chǎn)物的量直接計算出起始DNA拷貝數(shù)。<br /><br /><br type="_moz" /></span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　<strong>熒光定量PCR</strong>的方法：</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　熒光定量PCR所使用的熒光化學(xué)可分為兩種：熒光探針和熒光染料。而熒光定量 PCR 的方法相應(yīng)的可分為特異類和非特異類兩類，特異性檢測方法是在PCR反應(yīng)中利用標記熒光染料的基因特異寡核苷酸探針來檢測產(chǎn)物；而非特異性檢測方法是在在PCR反應(yīng)體系中，加入過量熒光染料，熒光染料特異性地摻入DNA雙鏈后，發(fā)射出熒光信號。前者由于增加了探針的識別步驟，特異性更高，但后者則簡便易行。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　<strong>熒光定量PCR</strong>zui常用的方法是DNA結(jié)合染料SYBR GreenⅠ的非特異性方法和Taqman水解探針的特異性方法，在這里主要介紹這2種方法，其它方法請參考其它熒光定量PCR資料。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　1、非特異性SYBR Green I染料法</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　SYBR Green I是一種結(jié)合于所有dsDNA雙螺旋小溝區(qū)域的具有綠色激發(fā)波長的染料（結(jié)合示意圖），在游離狀態(tài)下，SYBR Green I發(fā)出微弱的熒光，但一旦與雙鏈DNA結(jié)合后，熒光大大增強（作用機理圖）。因此，SYBR Green I的熒光信號強度與雙鏈DNA的數(shù)量相關(guān)，可以根據(jù)熒光信號檢測出PCR體系存在的雙鏈DNA數(shù)量。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　2、特異性Taqman水解探針法</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　Taqman熒光定量技術(shù)是以Taqman熒光探針為基礎(chǔ)，Taqman熒光探針為一寡核苷酸，兩端分別標記一個熒光發(fā)射基團和一個熒光淬滅基團。探針完整時，發(fā)射基團發(fā)射的熒光信號被淬滅基團吸收；</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　PCR擴增時，Taq酶的5’－3’外切酶活性將探針酶切降解，使熒光發(fā)射基團和熒光淬滅基團分離，從而熒光監(jiān)測系統(tǒng)可接收到熒光信號，即每擴增一條DNA鏈，就有一個熒光分子形成，實現(xiàn)了熒光信號的累積與PCR產(chǎn)物形成*同步。從而實現(xiàn)定量（如下圖）。常用的熒光基團是FAM, TET, VIC, HEX。</span></span></span></div></div> <div class="x4vfcpe" id="nr_textbox"><link rel="stylesheet" type="text/css" href="/css/MessageBoard_style.css"> <script language="javascript" src="/skins/Scripts/order.js?v=20210318" type="text/javascript"></script> <a name="order" id="order"></a> <div id="x4vfcpe" class="ly_msg" id="ly_msg"> <form 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