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NGS Library Prep kits

Molecular Research

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RNA isolater Total RNA Extraction Reagent

RNA isolater Total RNA Extraction Reagent, based on guanidine isothiocyanate and phenol, has a strong lysis ability, which can lyse cells and tissue samples in a short time to ensure the integrity of RNA. This product is widely used for culturing cells, animal tissues, microorganisms, and plant tissues with less secondary metabolism, such as seedlings and young leaves. The sample can be fully lysed in RNA isolater. After centrifugation with chloroform, the solution will form a supernatant layer, an intermediate layer and an organic layer (bright red lower layer). RNA is distributed in the upper aqueous phase. After the supernatant layer is collected, the Total RNA can be recovered by propanol precipitation.   Based on guanidine isothiocyanate and phenol, strong cracking ability; The optimized lysate formula has a wide range of sample adaptability.    Store at 4℃ and avoid light
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Number of views:
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Product serial number
R401-01
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RNA isolater Total RNA Extraction Reagent, based on guanidine isothiocyanate and phenol, has a strong lysis ability, which can lyse cells and tissue samples in a short time to ensure the integrity of RNA. This product is widely used for culturing cells, animal tissues, microorganisms, and plant tissues with less secondary metabolism, such as seedlings and young leaves. The sample can be fully lysed in RNA isolater. After centrifugation with chloroform, the solution will form a supernatant layer, an intermediate layer and an organic layer (bright red lower layer). RNA is distributed in the upper aqueous phase. After the supernatant layer is collected, the Total RNA can be recovered by propanol precipitation.

 

Based on guanidine isothiocyanate and phenol, strong cracking ability;
The optimized lysate formula has a wide range of sample adaptability. 

 

Store at 4℃ and avoid light

We could not find any corresponding parameters, please add them to the properties table

Q1:RNA降解。

A1:(1) 组织样本保存不当。RNA提取时要选新鲜组织或细胞样本,若为冻存样本,尽量避免反复冻融。样品离开活体或原来的生长环境后,样品中的内源RNase开始降解RNA,降解速度与内源RNase的含量及温度有关。

(2) 投入样本较多,导致裂解不充分。试剂不能有效抑制样品中所有的RNase。

(3) RNA保存时间过久,发生降解。对提取的RNA进行纯度和完整度检测,分管在-80℃长期保存或在-20℃短期保存,尽快使用,避免反复冻融。

Q2:组织样本应该如何保存。

A2:方法一:组织离体后迅速投入液氮冷冻,并用液氮保存。或待冻结后,将样本转移至 -80℃保存。注意:不能直接将新鲜组织直接放入 -80℃冷冻,否则样品的冻结会是一个缓慢的过程,在这个过程中内源 RNase 足以将 RNA 降解。

方法二:将新鲜组织充分浸泡于RNA Keeper® Tissue Stabilizer(R501)中,可在室温存放一周,4℃存放一个月,-20℃/-80℃长期保存。

Q3:提取的RNA有基因组DNA污染。

A3:(1) 向裂解液中加入氯仿后,需要在低温下(2-8℃)高速离心。离心后,RNA被抽提到上层的水相中,中、下层是有机相,含有氯仿,DNA即存在于中层。氯仿在常温下会与水以一定比例互溶,因此,常温离心会导致上层水相中也有少量基因组DNA的污染。吸取上层液体时,应非常小心,避免吸到中间层和下层。

(2) RNA样品中的少量基因组DNA残留,也可以通过后续逆转录反应前的基因组去除步骤进行去除,gDNA wiper能够高效去除100 ng基因组DNA。

Q4:加入异丙醇离心后无沉淀。

A4:RNA含量可能较低。建议加入异丙醇后在4℃或-20℃放置10-30 min后再离心。若依然看不见沉淀,在后面弃上清的操作时,采用吸取而不是倾倒的方法,以免沉淀丢失。

Q5:能否用于原核生物RNA的提取。

A5:可以,需配合R402(Bacteria RNA Plus Reagent)同时使用,可大幅提高原核生物RNA提取产量,同时保证良好的产物质量。

Q6:细胞样本比较少,该如何操作。

A6:裂解的过程中需要反复吹打细胞确保与裂解液充分混匀,冰上静置10 min。加入氯仿后需要剧烈振荡15s(难裂解的样品涡旋15s),然后在冰上静置5-10 min后离心,后续就常规沉淀提取。

Q7:液氮研磨组织时,若组织变为冻霜状,比较粘连,该如何操作。

A7:将组织液氮研磨成粉末后,转移至提前在液氮中预冷的EP管中,加入RNA isolater后用枪上下吹直至将絮状物吹散后,进行后续实验。同时可以减小研磨组织的体积,大小等同于指甲盖一半的体积,减少絮状物的产生。

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