|RNA Recovery||100 µg||50 µg||10 µg|
|Minimum Elution||50 µl||25 µl||6 µl|
|(Animal) Cells||≤ 107||≤ 5 x 106||≤ 106|
|Tissue||≤ 50 mg||≤ 25 mg||≤ 5 mg|
The Direct-zol RNA Kits provide a streamlined method for the purification of up to 100 µg (per prep) of high-quality RNA directly from samples in TRI Reagent or similar. Total RNA, including small RNAs (17-200 nt), is effectively isolated from a variety of sample sources (cells, tissues, serum, plasma, blood, biological liquids, etc.).
Isolation of RNA by conventional phase separation was shown to selectively enrich for some species of miRNA, leading to bias in downstream analysis. The Direct-zol method assures unbiased recovery of small RNAs including miRNA (see below).
The procedure is easy. Simply apply a prepared sample in TRI Reagent directly to the Zymo-Spin Column and then bind, wash, and elute the RNA. No phase separation, precipitation, or post-purification steps are necessary. The eluted RNA is high quality and suitable for subsequent molecular manipulation and analysis (including RT-PCR, transcription profiling, hybridization, sequencing etc.).
The entire procedure typically takes only 7 minutes.
|Sample Sources||Any sample stored and preserved in TRI Reagent®, TRIzol® or similar. (animal cells, tissue, bacteria, yeast, biological fluids, and in vitro processed RNA (e.g., transcription products, DNase-treated or labeled RNA)).|
|RNA Purity||A260/A280 >1.8, A260/A230 >1.8. Complete removal of DNA can be accomplished using an in-column DNase I digestion|
|Compatibility||TRIzol®, RNAzol®, QIAzol®, TriPure™, TriSure™ and all other acid-guanidinium-phenol based solutions can be used in place of TRI Reagent®.|
|RNA Storage||RNA eluted with DNase/RNase-Free Water (provided) can be stored at ≤-70 ºC. The addition of RNase inhibitors is highly recommended for prolonged storage.|
|RNA Size Limits||RNAs ≥17 nucleotides.|
|Sample Inactivation||TRI Reagent® (provided with R2051, R2053, R2061, R2063, R2071, R2073 only) inhibits RNase activity and inactivates viruses and other infectious agents.|
High quality broad range RNA is purified with the Direct-zol™ RNA MiniPrep. (A) DNA-free RNA purified from human epithelial cells using the Direct-zol™ RNA MiniPrep compared to a DNA containing preparation from Supplier Q (1% agarose/TAE). (B) Small RNAs are effectively recovered with the Direct-zol™ procedure while absent in Supplier Q preparations
(Agilent Bioanalyzer 2100, Small RNA Chip data shown)
|The data show RNA purified from TRIzol® samples using the Direct-zol™ RNA MiniPrep compared to an unbiased method (mirVana™, Ambion). Micro-RNA analysis was performed using miRNA-Seq (MiSeq®, Illumina) and a direct hybridization assay (nCounter®, Nanostring).|