RN38-EASYspin Plus 植物RNA快速提取试剂盒
货号# | 规格 | 价格 |
RN3802 | 50 次 | 1300.00 元 |
产品介绍:
本公司推出EASYspin无苯酚、氯仿RNA快速提取技术基础上,又成功研发基因组DNA清除柱技术可以有效清除gDNA残留,得到的RNA一般不需要DNase消化,可用于反转录PCR、荧光定量PCR等实验。独特的裂解液迅速裂解细胞和灭活细胞RNA酶,植物RNA助提剂PLANTaid帮助结合多糖多酚并通过离心去除,然后裂解混合物用乙醇调节RNA结合吸附到基因组DNA清除柱,基因组DNA清除柱子同时吸附清除残留的DNA, 然后RNA被选择性洗脱滤过。滤过的RNA用乙醇调节结合条件后,RNA在高离序盐状态下选择性吸附于离心柱内硅基质膜, 再通过一系列快速的漂洗-离心的步骤, 去蛋白液和漂洗液将细胞代谢物,蛋白等杂质去除, 最后低盐的RNase free H20将纯净RNA从硅基质膜上洗脱。
产品特点:
1.完全不使用有毒的苯酚,氯仿等试剂,也不需要乙醇沉淀等步骤。
2.简捷,单个样品操作一般可在25分钟内完成,世界上最简单快速的试剂盒。
3.独有的植物RNA助提剂可以有效结合多糖多酚,提高清除效果。
4.独家研发成功基因组DNA清除柱技术可以有效清除gDNA残留,得到的RNA一般不需要DNase消化,可用于反转录PCR、荧光定量PCR等实验。
5.适应性极其广泛,可以提取包括棉花、松针、冬青树叶、葡萄叶片、等100多种国内外试剂盒提取失败的样品。详细样品列表请参考公司主页产品介绍。
6.多次柱漂洗确保高纯度,OD260/OD280典型的比值达1.9~2.2,基本无DNA残留,可用于RT-PCR,Northern-blot和各种实验。
使用RN38-EASYspin Plus 植物RNA快速提取试剂盒 发表文献
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其它公司多糖多酚植物RNA提取试剂盒失败原因和解决方案
很多植物RNA的样品由于含有大量的多糖、多酚、代谢产物、色素等成分,造成RNA提取过程中氧化、褐化、降解、由于植物品种的多样性造成情况更加复杂。手工的CTAB类的方法提取因为时间太长,太繁琐,手工方法不在讨论之列。一直以来没有一款好的试剂盒包括qiagen、promega等进口试剂盒也无法满足科研工作者对植物RNA提取的要求。
下面我们来分析一下植物RNA为什么不能提取成功的原因:
市面上最常见的RNA提取试剂盒无非是两种:种:TRIzol改良类方法(包括溶液型的和离心柱型的)、第二种:直接裂解过柱子的方法(离心柱)
种试剂盒失败的原因1:RNA市面上面最流行的方法就是Trizol,或者Triol改良,或者Trizol加离心柱一类的改良方法。trizol也就是异硫氰酸胍/苯酚/氯仿原理一步法的方法最适合的对象是动物源性的组织细胞,针对普通多糖多酚低的植物性的材料,TRIzol类原理产品也可以提取。但是多糖、多酚、次级代谢产物丰富的情况下,trizol类方法无法防止多糖多酚对于RNA/DNA分相的干扰,要么残留大量DNA,要么残留大量多糖、多酚或者次级代谢产物,氧化破坏RNA,或者残留这些多糖多酚,色素代谢产物等抑制下游的反转录等反应。限于技术水平的限制,市面上绝大多数的国产厂家是使用trizol的方法进行改良,无论是不是加了离心柱。但是实践证明,改良不能从根本上解决问题。判断是否试剂盒使用这种改良的方法非常简单: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。
第二种试剂盒失败的原因:直接裂解过柱子的方法是目前最先进的方法,但是也是技术含量最高的方法。这个方法采用裂解液(不含苯酚,氯仿)直接裂解,RNA/DNA同时过柱子,然后在柱子上面直接分离RNA/DNA,所以,这种方法的优点在于,避免了使用trizol在多糖多酚下不能成功分离RNA/DNA的弊端、第二在于,不使用有毒的苯酚氯仿。但是正是因为其技术先进,所以难度很高,国内厂家包括进口公司有两个技术难点一直没有突破。,裂解液的成分必须针对去除多糖多酚进行研发添加去多糖多酚,代谢产物成分。否则会同样碰到多糖多酚干扰提取的问题。第二、和trizol原理不同,直接过柱法DNA/RNA同时加到吸附柱上去。如何去除DNA是个难点。否则会残留大量DNA。两个技术难点的没有掌握导致了国内公司包括的第二种试剂盒失败。国外公司因为没有掌握难点,裂解液里面没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样品。
部分成功样品:
植物:棉花、海棠、黑加仑、烟草、拟南芥、虎杖、大豆、草莓、冬青、月季花雌蕊、蔷薇、沙棘、冬枣、芦荟、仙人掌、报春花、水稻、玉米、唐菖蒲、樱桃、白玉兰、毛白杨、樱花、葡萄、百合花、百合叶子雌蕊雄蕊、紫菜、绿藻、香蕉、水仙花、青花菜、地被菊、苹果、梅花、番茄、石斛、毛桃、苎麻、慈姑、葛根、甘肃桃、玫瑰花、槟榔果、甜糖菊、硅藻、牡丹、胡杨、油桐果、梨子皮、板栗花序、青皮云杉、红树根、铁线蕨、黄瓜、小麦叶子种子、番木瓜、甘薯、紫薯、油松、油茶、马尾松、芜菁、毛果杨、木薯、大叶落地生根、山杏、旱柳、桉树、琵琶花果、丹参、人参、西洋参、栀子、洋葱、红豆杉、梨树叶、五倍子、泡桐、西瓜、芍药、雪莲等等,其中包括qiagen无法提取的黑加仑、冬青、月季、松针、葡萄叶片等,promega无法提取的海棠等样品、均可用该产品成功提取。