TRIeasy™ LS液体总RNA提取试剂(同TRIzol LS)|TRIeasy™ LS Total RNA Extraction Reagent

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TRIeasy™ LS液体总RNA提取试剂(同TRIzol LS)|TRIeasy™ LS Total RNA Extraction Reagent

产品说明书

FAQ

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已发表文献

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。

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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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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

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Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus)

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Hifair® qPCR SYBR® Green Master Mix (No Rox )

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Hifair® qPCR SYBR® Green Master Mix (Low Rox Plus) 

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Hifair® qPCR SYBR® Green Master Mix (High Rox Plus) 

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HB210713

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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