RN38-EASYspin Plus 植物RNA快速提取試劑盒

產(chǎn)品介紹：

     本公司推出EASYspin無(wú)苯酚、氯仿RNA快速提取技術(shù)基礎(chǔ)上，又成功研發(fā)基因組DNA清除柱技術(shù)可以有效清除gDNA殘留，得到的RNA一般不需要DNase消化，可用于反轉(zhuǎn)錄PCR、熒光定量PCR等實(shí)驗(yàn)。獨(dú)特的裂解液迅速裂解細(xì)胞和滅活細(xì)胞RNA酶，植物RNA助提劑PLANTaid幫助結(jié)合多糖多酚并通過(guò)離心去除，然后裂解混合物用乙醇調(diào)節(jié)RNA結(jié)合吸附到基因組DNA清除柱，基因組DNA清除柱子同時(shí)吸附清除殘留的DNA, 然后RNA被選擇性洗脫濾過(guò)。濾過(guò)的RNA用乙醇調(diào)節(jié)結(jié)合條件后，RNA在高離序鹽狀態(tài)下選擇性吸附于離心柱內(nèi)硅基質(zhì)膜， 再通過(guò)一系列快速的漂洗－離心的步驟, 去蛋白液和漂洗液將細(xì)胞代謝物，蛋白等雜質(zhì)去除， 最后低鹽的RNase free H20將純凈RNA從硅基質(zhì)膜上洗脫。

產(chǎn)品特點(diǎn)：

1.完全不使用有毒的苯酚，氯仿等試劑，也不需要乙醇沉淀等步驟。 

2.簡(jiǎn)捷，單個(gè)樣品操作一般可在25分鐘內(nèi)完成，世界上最簡(jiǎn)單快速的試劑盒。 

3.獨(dú)有的植物RNA助提劑可以有效結(jié)合多糖多酚，提高清除效果。 

4.獨(dú)家研發(fā)成功基因組DNA清除柱技術(shù)可以有效清除gDNA殘留，得到的RNA一般不需要DNase消化，可用于反轉(zhuǎn)錄PCR、熒光定量PCR等實(shí)驗(yàn)。 

5.適應(yīng)性極其廣泛，可以提取包括棉花、松針、冬青樹葉、葡萄葉片、等100多種國(guó)內(nèi)外試劑盒提取失敗的樣品。詳細(xì)樣品列表請(qǐng)參考公司主頁(yè)產(chǎn)品介紹。 

6.多次柱漂洗確保高純度，OD260/OD280典型的比值達(dá)1.9～2.2，基本無(wú)DNA殘留,可用于RT-PCR，Northern-blot和各種實(shí)驗(yàn)。

使用RN38-EASYspin Plus 植物RNA快速提取試劑盒 發(fā)表文獻(xiàn)

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其它公司多糖多酚植物RNA提取試劑盒失敗原因和解決方案

很多植物RNA的樣品由于含有大量的多糖、多酚、代謝產(chǎn)物、色素等成分，造成RNA提取過(guò)程中氧化、褐化、降解、由于植物品種的多樣性造成情況更加復(fù)雜。手工的CTAB類的方法提取因?yàn)闀r(shí)間太長(zhǎng)，太繁瑣，手工方法不在討論之列。一直以來(lái)沒(méi)有一款好的試劑盒包括qiagen、promega等進(jìn)口試劑盒也無(wú)法滿足科研工作者對(duì)植物RNA提取的要求。

下面我們來(lái)分析一下植物RNA為什么不能提取成功的原因：

市面上最常見的RNA提取試劑盒無(wú)非是兩種：種：TRIzol改良類方法（包括溶液型的和離心柱型的）、第二種：直接裂解過(guò)柱子的方法（離心柱）

種試劑盒失敗的原因1：RNA市面上面最流行的方法就是Trizol，或者Triol改良，或者Trizol加離心柱一類的改良方法。trizol也就是異硫氰酸胍/苯酚/氯仿原理一步法的方法最適合的對(duì)象是動(dòng)物源性的組織細(xì)胞，針對(duì)普通多糖多酚低的植物性的材料，TRIzol類原理產(chǎn)品也可以提取。但是多糖、多酚、次級(jí)代謝產(chǎn)物豐富的情況下，trizol類方法無(wú)法防止多糖多酚對(duì)于RNA/DNA分相的干擾，要么殘留大量DNA，要么殘留大量多糖、多酚或者次級(jí)代謝產(chǎn)物，氧化破壞RNA，或者殘留這些多糖多酚，色素代謝產(chǎn)物等抑制下游的反轉(zhuǎn)錄等反應(yīng)。限于技術(shù)水平的限制，市面上絕大多數(shù)的國(guó)產(chǎn)廠家是使用trizol的方法進(jìn)行改良，無(wú)論是不是加了離心柱。但是實(shí)踐證明，改良不能從根本上解決問(wèn)題。判斷是否試劑盒使用這種改良的方法非常簡(jiǎn)單：是否裂解液含有苯酚的味道和使用氯仿，如果使用到了氯仿就是TRIzol方法的改良。

第二種試劑盒失敗的原因：直接裂解過(guò)柱子的方法是目前最先進(jìn)的方法，但是也是技術(shù)含量最高的方法。這個(gè)方法采用裂解液（不含苯酚，氯仿）直接裂解，RNA/DNA同時(shí)過(guò)柱子，然后在柱子上面直接分離RNA/DNA，所以，這種方法的優(yōu)點(diǎn)在于，避免了使用trizol在多糖多酚下不能成功分離RNA/DNA的弊端、第二在于，不使用有毒的苯酚氯仿。但是正是因?yàn)槠浼夹g(shù)先進(jìn)，所以難度很高，國(guó)內(nèi)廠家包括進(jìn)口公司有兩個(gè)技術(shù)難點(diǎn)一直沒(méi)有突破。，裂解液的成分必須針對(duì)去除多糖多酚進(jìn)行研發(fā)添加去多糖多酚，代謝產(chǎn)物成分。否則會(huì)同樣碰到多糖多酚干擾提取的問(wèn)題。第二、和trizol原理不同，直接過(guò)柱法DNA/RNA同時(shí)加到吸附柱上去。如何去除DNA是個(gè)難點(diǎn)。否則會(huì)殘留大量DNA。兩個(gè)技術(shù)難點(diǎn)的沒(méi)有掌握導(dǎo)致了國(guó)內(nèi)公司包括的第二種試劑盒失敗。國(guó)外公司因?yàn)闆](méi)有掌握難點(diǎn)，裂解液里面沒(méi)有去除多糖多酚成分，所以包括qiagen的盒子也常常不能成功提取植物RNA樣品。

部分成功樣品：

植物：棉花、海棠、黑加侖、煙草、擬南芥、虎杖、大豆、草莓、冬青、月季花雌蕊、薔薇、沙棘、冬棗、蘆薈、仙人掌、報(bào)春花、水稻、玉米、唐菖蒲、櫻桃、白玉蘭、毛白楊、櫻花、葡萄、百合花、百合葉子雌蕊雄蕊、紫菜、綠藻、香蕉、水仙花、青花菜、地被菊、蘋果、梅花、番茄、石斛、毛桃、苧麻、慈姑、葛根、甘肅桃、玫瑰花、檳榔果、甜糖菊、硅藻、牡丹、胡楊、油桐果、梨子皮、板栗花序、青皮云杉、紅樹根、鐵線蕨、黃瓜、小麥葉子種子、番木瓜、甘薯、紫薯、油松、油茶、馬尾松、蕪菁、毛果楊、木薯、大葉落地生根、山杏、旱柳、桉樹、琵琶花果、丹參、人參、西洋參、梔子、洋蔥、紅豆杉、梨樹葉、五倍子、泡桐、西瓜、芍藥、雪蓮等等，其中包括qiagen無(wú)法提取的黑加侖、冬青、月季、松針、葡萄葉片等，promega無(wú)法提取的海棠等樣品、均可用該產(chǎn)品成功提取。