About Direct-zol™ RNA MiniPrep
The Direct-zol™ RNA MiniPrep provides a streamlined method for the purification of up to 50 µg (per prep) of high-quality RNA directly from samples in TRI-Reagent® or similar*. Total RNA (including small and non-coding RNAs) is effectively isolated from a variety of sample sources (cells, tissues, biological liquids, etc.) using this product.
The procedure is easy: simply apply a sample in TRI-Reagent® to spin column and then spin, wash, and elute the RNA. No phase separation, precipitation, or post-purification steps are necessary. Small RNAs (≥17 nucleotides) are efficiently and consistently recovered using this kit. The result is broad range purification of small and large RNAs suitable for subsequent RNA-based methods including RT-PCR, transcription profiling, hybridization, etc.
The entire procedure typically takes about 5-10 minutes.
| Format | Spin Column |
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| Processing Time | 10 min |
| Equipment | Microcentrifuge |
| Sample Sources | Cells from culture, solid tissue, plasma, serum, whole blood, and in vitro processed RNA (e.g., transcription products, DNase-treated or labeled RNA) or samples stored and preserved in TRI-Reagent®, TRIzol®, or similar. |
| RNA Purity | High quality RNA (A260/A280 >1.8, A260/A230 >1.8) is recovered. |
| RNA Recovery | The RNA binding capacity of the supplied Zymo-Spin™ IIC Column is ~50 µg with a minimal elution volume of 25 µl. |
| Compatibility | TRI-Reagent®, TRIzol®, RNAzol®, QIAzol®, TriPure, RNA-Bee or similar acid-guanidinium-phenol based solutions. |
| RNA Storage | ≤-70 ºC. The addition of RNase inhibitors (optional) is highly recommended for prolonged storage. |
| RNA Size Limits | RNAs ≥17 nucleotides. |
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| 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) | Viral RNA is detected with high sensitivity following the Direct-zol™ isolation method. The Direct-zol™ method significantly improves the detection of West Nile virus when compared to the conventional phaseseparation method. The RT-qPCR data show ?Ct = 5 (average of two independent experiments). RNA was isolated from cell-free samples inactivated using the TRIReagent®. |
| Step 1 | Add one volume ethanol (95-100%) directly to one volume sample homogenate (1:1) in TRI Reagent® or similar1. Mix well by vortexing. |
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| Step 2 | Load the mixture into a Zymo-Spin™ IIC Column2 in a Collection Tube and centrifuge for 1 minute. Transfer the column into a new Collection Tube and discard the Collection Tube containing the flow-through. Note: At this point, RNA samples can be in-column DNase treated. |
| Step 3 | Add 400 µl Direct-zol™ RNA PreWash3 to the column and centrifuge for 1 minute. Discard the flow-through. Repeat this step. Add 700 µl RNA Wash Buffer3 to the column and centrifuge for 1 minute. Discard the flow-through. To ensure complete removal of the wash buffer, centrifuge the column for an additional 2 minutes in an emptied Collection Tube. Transfer the column carefully into an RNase-free tube (not provided). |
| Step 4 | Add ≥25 µl of DNase/RNase-Free Water4 directly to the column matrix and centrifuge at max speed for 1 minute. The eluted RNA can be used immediately or stored at ≤-70°C. |
Featured Citations
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Total RNA was extracted from human, rat, and mouse cortical neurons using the Direct-zol™ RNA MiniPrep Kit. The high-quality RNA was reverse transcribed into cDNA and used to investigate the effect of BPA exposure on neurodevelopment by real-time PCR analysis.
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High-quality RNA isolated with the Direct-zol™ RNA MiniPrep from fecal and culture samples were shown to be instrumental in the development of a rotavirus early detection system using RT-PCR. The efficient RNA isolation helps in identification of severe gastroenteritis in infants and young children.
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The Direct-zol™ RNA MiniPrep isolated RNA from Vibrio cholerae has been used for next-gen sequencing, ChIP-Seq, RNA-Seq, qPCR and Northern-blot analysis. The high-quality RNA helped to characterize gene expression profiles of virulence factors in RpoN regulon of the cholera pathogen.
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The Direct-zol RNA Miniprep kit was used to isolate pure total RNA from Rat cells, and the high quality RNA was used to create a cDNA library for RT-qPCR analysis of several genes related to hematopoietic Stem Cell (HSC) Development. The consistent RNA extraction allowed the scientists to determine that HSC cells do not significantly contribute to kidney repair following acute kidney injuries.
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“I am very impressed with the RNA yield (quality and quantity) I obtained using this kit. When using other methods and kits for RNA isolation- despite us taking several steps and measures to reduce RNA degradation- I found it very challenging to obtain good quality RNA. With Direct-zol RNA mini prep I was able to obtain superior quality RNA repeatedly and with ease. This kit is suitable for both small scale and high throughput RNA isolations.”
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Subashini N. (Wayne State University)
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“The protocol very straight forward, time needed to get the RNA is just outstanding and saved us a lot of time to do more things. The yield and the purity of the RNA is something out of this world, about at least 50% more RNA and purity not less than 1.95, we never got [that] before with other vendors such Invitrogen, Qiagen, etc.”
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Ehab S. (University of Iowa)
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“I liked that it wasn't necessary to separate phases with Chloroform - I think I got less gDNA contamination as a result. The option for performing an on-column DNAse1 digestion is perhaps the most significant benefit compared to doing ChCl3 separations and subsequent precipitations”
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Nick A. (AgResearch, New Zealand)
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“It is very easy to use and centrifugations are short. The resin is compact and absorbent which makes it easy to cover the whole resin surface with small amount of water in the elution step. Recovery of small RNAs is very good based on CT values obtained from qPCR using RNA template derived from this kit.”
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Janne T. (Universidad de Zaragoza)
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“The speed of the process is by far the best aspect of this kit. Previously I used a protocol that took 3 hours, now I can have my RNA in 20 minutes. What is not to like about that? What I also really liked is the ease of the whole process: no messing around with several buffers or different columns that get switched up. Just one column and two buffers, I love it. Oh, and the RNA yield was great too.”
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Arjan V. (Indiana University)
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“No phase separation was needed, but you still had the benefits of a Trizol extraction. No need to precipitate and resuspend samples, which means less sample loss during purification.”
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Adina B. (University of Guelph)
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“Simple protocol and yielded good quality of RNA. Only one kit working for all type of tissue, cell and especially biological fluids.”
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Mohan K. (University of Illinois, Chicago)
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“Easy to use, equal results as a much more expensive kit from another company.”
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Patricia O. (University of Porto)
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"I've been performing RNA isolation on dozens of samples using the manual method, and this kit is amazing. It really is true to its prep time: 10 minutes. It's so much easier, and given all the other experimental methods I need to perform, this miniprep allows me to run more samples than ever before! I've got a gel comparing the lack of gDNA as shown in the advertising pamphlet. What can I say, except: I love this product!"
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R.K. CSU
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