qiagen 74136DNA微量提取試劑盒

操作流程

RNeasy Plus Micro Kit能從多達(dá)5 x 105個細(xì)胞或5 mg組織中分離總RNA，RNeasy Plus Mini Kit能夠從107個細(xì)胞或30 mg組織中分離總RNA。簡潔的工作流程可在25分鐘內(nèi)純化得到已去除基因組DNA的RNA。樣本首先被裂解和勻質(zhì)化。裂解液流過gDNA Eliminator柱，在流過gDNA Eliminator柱的溶液中加入乙醇，樣本再上樣到RNeasy離心柱。RNA結(jié)合到膜上，污染物被洗去。獲得高品質(zhì)RNA，RNeasy Plus Micro Kit的洗脫體積為14 µl，RNeasy Plus Mini Kit的洗脫體積為30 µl。

處理不同的起始樣本可用不同的實驗方案。實驗方案的不同之處主要在于樣本的裂解和勻質(zhì)化。樣本上樣到gDNA Eliminator柱之后的步驟就類似。

使用Buffer RLT Plus（RNeasy Plus Micro Kit提供）進行組織破碎和勻質(zhì)化時，可能會產(chǎn)生過多泡沫。在破碎和勻漿前向Buffer RLT Plus中加入Reagent DX（單獨提供）可充分減少泡沫產(chǎn)生。Reagent DX經(jīng)仔細(xì)檢測，表明配合此試劑盒使用不會影響RNA的純度或下游應(yīng)用。

qiagen 74136DNA微量提取試劑盒

For purification of up to 100 µg total RNA from cells/tissues using gDNA Eliminator columns

• Unique gDNA Eliminator columns avoid the need for DNase
• Efficient removal of genomic DNA
• Highly reproducible yields of RNA in minutes
• High-performance RNA for sensitive applications

The RNeasy Plus Mini Kit integrates fast, convenient purification of up to 100 µg RNA with effective elimination of genomic DNA. Cell or tissue lysates are spun through gDNA Eliminator spin columns to remove genomic DNA. Total RNA is then purified using RNeasy Mini spin columns. The kit can be automated on theQIAcube. Tissue samples can be conveniently stabilized using RNAlater RNA Stabilization Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. For smaller samples, the RNeasy Plus Micro Kit (spin-column binding capacity of 45 µg RNA) is also available.

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