產(chǎn)品說明書
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已發(fā)表文獻
TRIeasyTM LS Total RNA Extraction Reagent 是一種適用于各種動植物、酵母、細菌等液體樣本的總RNA抽提試劑，具有極強的裂解能力，可在短時間內(nèi)裂解細胞和組織樣本，并有效抑制樣本中RNA的降解，保持RNA的完整性。樣品在該試劑中能夠充分裂解，之后加入氯仿離心分層，形成上清層、中間層和有機層，RNA分布在上層水相中，收集上清層后，經(jīng)異丙醇沉淀便可得到總RNA。提取的總RNA純度高，基本不含蛋白質(zhì)及基因組DNA，可直接用于Northern、點雜交、mRNA純化、體外翻譯、RT-PCR、poly(A)+選擇、RNA酶保護分析以及構(gòu)建cDNA文庫等多種分子生物學實驗。
本品操作簡單快速，所有操作可在一小時內(nèi)完成。對少量的組織(50-100 mg)和細胞(5×106)以及大量的組織(≥1 g)和細胞(>107)均有較好的裂解效果。
運輸和保存方法
冰袋運輸。4℃避光保存，有效期一年。
相關(guān)產(chǎn)品
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產(chǎn)品名稱 |
貨號 |
規(guī)格 |
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Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) |
11141ES60 |
100 T |
|
11201ES08 |
5 mL |
|
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11202ES08 |
5 mL |
|
|
11203ES08 |
5 mL |
HB210713
Q：10606和19201的區(qū)別是什么？
A：19201是濃縮形式，需要的試劑用量更少一點，提取的準確性和提取反應的速度更快一些。一般10606就可以滿足常規(guī)提取需求，如果客戶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) 組織樣本保存不當。RNA 提取時要選新鮮組織或細胞樣本，若為凍存樣本，盡量避免反復凍融。樣品離開活體或原來的生長環(huán)境后，樣品中的內(nèi)源RNase 開始降解 RNA，降解速度與內(nèi)源 RNase 的含量及溫度有關(guān)；(2) 投入樣本較多，導致裂解不充分。試劑不能有效抑制樣品中所有的 RNase；(3) RNA 保存時間過久，發(fā)生降解。對提取的RNA 進行純度和完整度檢測，分管在-80℃長期保存或在-20℃短期保存，盡快使用，避免反復凍融。
Q:是否有提 RNA 的層析柱?
A:沒有。
Q:提取的 RNA 有基因組 DNA 污染？
A:(1) 向裂解液中加入氯仿后，需要在低溫下（2-8℃）高速離心。離心后，RNA 被抽提到上層的水相中，中、下層是有機相，含有氯仿，DNA 即存在于中層。氯仿在常溫下會與水以一定比例互溶，因此，常溫離心會導致上層水相中也有少量基因組DNA 的污染。吸取上層液體時， 應非常小心，避免吸到中間層和下層；(2) RNA 樣品中的少量基因組 DNA 殘留，也可以通過后續(xù)逆轉(zhuǎn)錄反應前的基因組去除步驟進行去除。
Q:加入異丙醇離心后無沉淀？
A:RNA 含量可能較低。建議加入異丙醇后在 4℃或-20℃放置 10-30 min 后再離心。若依然看不見沉淀，在后面棄上清的操作時，采用吸取而不是傾倒的方法，以免沉淀丟失。
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