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class="jyms"><p><span>簡(jiǎn)要描述：</span>RNA可分為信使RNA、轉(zhuǎn)運(yùn)RNA和核糖體RNA三大類，不同組織總RNA提取實(shí)質(zhì)就是將細(xì)胞裂解，釋放出RNA，并通過(guò)不同方式去除蛋白、DNA等雜質(zhì)，Z終獲得高純RNA產(chǎn)物的過(guò)程。由于RNA樣品易受環(huán)境因素特別是RNA酶的影響而降解，提取高質(zhì)量的RNA樣品在生命科研中具有相當(dāng)?shù)奶魬?zhàn)性。RNA提取對(duì)樣品的新鮮性要求非常高，獲取樣品后立即提取RNA，若無(wú)條件立即實(shí)驗(yàn)，應(yīng)于-80℃或液氮中。</p></div> <ul> <li><img src="/Skins/270236/images/dot.png" /><span>產(chǎn)品型號(hào)：</span></li> <li><img src="/Skins/270236/images/dot.png" /><span>廠商性質(zhì)：</span>生產(chǎn)廠家</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>訪問(wèn) 量：</span>2947</li> </ul> <div id="priymv99ks" 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="priymv99ks" class="clear"></div> </div>  <div class="priymv99ks" id="ny_pro_box02"> <div class="priymv99ks" id="con"> <ul id="tags"> <li id="priymv99ks" class=selectTag><A onmouseover="selectTag('tagContent0',this)" href="javascript:void(0)"onFocus="this.blur()">產(chǎn)品介紹：</A> </li> </ul> <div class="priymv99ks" id=tagContent> <div id="priymv99ks" class="tagContent selectTag" id=tagContent0><strong> <span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">RNA抽提</span></span></span></strong><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">操作步驟:</span></span></span><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　1. 勻漿處理:</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　a.組織將組織在液氮中磨碎,每50-100mg組織加入1ml TRIzol,用勻漿儀進(jìn)行勻漿處理。樣品體積不應(yīng)超過(guò)TRIzol體積10℅。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　b.單層培養(yǎng)細(xì)胞直接在培養(yǎng)板中加入TRIzol裂解細(xì)胞，每10cm2面積（即3.5cm直徑的培養(yǎng)板）加1ml，用移液器吸打幾次。TRIzol的用量應(yīng)根據(jù)培養(yǎng)板面積而定，不取決于細(xì)胞數(shù)。TRIzol加量不足可能導(dǎo)致提取的RNA有DNA污染。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　c．細(xì)胞懸液離心收集細(xì)胞，每5-10×106動(dòng)物、植物、酵母細(xì)胞或1×107細(xì)菌細(xì)胞加入1ml TRIzol，反復(fù)吸打。加TRIzol之前不要洗滌細(xì)胞以免mRNA降解。一些酵母和細(xì)菌細(xì)胞需用勻漿儀處理。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　2．將勻漿樣品在室溫（15-30℃）放置5分鐘，使核酸蛋白復(fù)合物*分離。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　3．可選步驟：如樣品中含有較多蛋白質(zhì)，脂肪，多糖或胞外物質(zhì)（肌肉，植物結(jié)節(jié)部分等）可于2-8℃10000×g離心10分鐘，取上清。離心得到的沉淀中包括細(xì)胞外膜，多糖，高分子量DNA，上清中含有RNA。處理脂肪組織時(shí)，上層有大量油脂應(yīng)去除。取澄清的勻漿液進(jìn)行下一步操作。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　5. 每使用1ml TRIzol加入0.2ml氯仿，劇烈振蕩15秒，室溫放置3分鐘。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　6. 2-8℃10000×g離心15分鐘。樣品分為三層：底層為黃色有機(jī)相，上層為無(wú)色水相和一個(gè)中間層。RNA主要在水相中，水相體積約為所用TRIzol試劑的60℅。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　7. 把水相轉(zhuǎn)移到新管中，如要分離DNA和蛋白質(zhì)可保留有機(jī)相，進(jìn)一步操作見(jiàn)后。用異丙醇沉淀水相中的RNA。每使用1ml TRIzol加入0.5ml異丙醇，室溫放置10分鐘。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　8. 2-8℃10000×g離心10分鐘，離心前看不出RNA沉淀，離心后在管側(cè)和管底出現(xiàn)膠狀沉淀。移去上清。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　9. 用75℅乙醇洗滌RNA沉淀。每使用1ml TRIzol至少加1ml 75℅乙醇。2-8℃不超過(guò)7500×g離心5分鐘，棄上清。</span></span></span></div><div><span style="color: rgb(0, 0, 0);"><span style="font-size: small;"><span style="font-family: Arial;">　　10.室溫放置干燥或真空抽干RNA沉淀，大約晾5-10分鐘即可.不要真空離心干燥，過(guò)于干燥會(huì)導(dǎo)致RNA的溶解性大大降低。加入25-200μl無(wú)RNase的水或0.5℅SDS,用槍頭吸打幾次,55-60℃放置10分鐘使RNA溶解.如RNA用于酶切反應(yīng),勿使用SDS溶液。RNA也可用100℅的去離子甲酰胺溶解，-70℃保存。</span></span></span></div></div> <div class="priymv99ks" 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="priymv99ks" class="ly_msg" id="ly_msg"> <form method="post" name="form2" id="form2"> <h3>留言框 </h3> <ul> <li> <h4 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