TRIeasyTM LS Total RNA Extraction Reagent 是一种适用于各种动植物、酵母、细菌等液体样本的总RNA抽提试剂,具有极强的裂解能力,可在短时间内裂解细胞和组织样本,并有效抑制样本中RNA的降解,保持RNA的完整性。样品在该试剂中能够充分裂解,之后加入氯仿离心分层,形成上清层、中间层和有机层,RNA分布在上层水相中,收集上清层后,经异丙醇沉淀便可得到总RNA。提取的总RNA纯度高,基本不含蛋白质及基因组DNA,可直接用于Northern、点杂交、mRNA纯化、体外翻译、RT-PCR、poly(A)+选择、RNA酶保护分析以及构建cDNA文库等多种分子生物学实验。
本品操作简单快速,所有操作可在一小时内完成。对少量的组织(50-100 mg)和细胞(5×106)以及大量的组织(≥1 g)和细胞(>107)均有较好的裂解效果。
运输和保存方法
冰袋运输。4℃避光保存,有效期一年。
相关产品
产品名称 |
货号 |
规格 |
Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) |
11141ES60 |
100 T |
11201ES08 |
5 mL |
|
11202ES08 |
5 mL |
|
11203ES08 |
5 mL |
HB210713
Q:10606和19201的区别是什么?
A:19201是浓缩形式,需要的试剂用量更少一点,提取的准确性和提取反应的速度更快一些。一般10606就可以满足常规提取需求,如果客户想好更好的提取效果和rnase抑制效果可以用19201。
Q:RNA 降解?
A:(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。
Q:是否有提 RNA 的层析柱?
A:没有。
Q:提取的 RNA 有基因组 DNA 污染?
A:(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。
Q:加入异丙醇离心后无沉淀?
A: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。
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