Highlights

• Easy Handling: Bypass chloroform, phase separation and precipitation steps.
• NGS-Ready: Ultra-pure RNA without phenol carryover. No DNA contamination (DNase I included).
• Non-Biased: Complete RNA recovery without miRNA loss.

Description

The Direct-zol™ RNA MiniPrep Plus Kits are RNA purification kits that 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. 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.

Learn More

Performance

Technical Specifications

Compatibility	TRIzol®, RNAzol®, QIAzol®, TriPure™, TriSure™ and all other acid-guanidinium-phenol based solutions can be used in place of TRI Reagent®.
Equipment	Microcentrifuge, vortex
Sample Inactivation	TRI Reagent® (provided with R2071, R2073) inhibits RNase activity and inactivates viruses and other infectious agents.
Sample Source	Any sample stored and preserved in TRI Reagent®, TRIzol® or similar (animal cells, tissue, bacteria, yeast, fecal, biological fluids, and in vitro processed RNA (e.g., transcription products, DNase-treated or labeled RNA)).
Size Range	Total RNA ≥ 17 nt
Yield	100 µg RNA (binding capacity), ≥ 50 µl (elution volume)

Resources

Documents

Protocol: 

Datasheet: 

SDS (MSDS): 

SDS (MSDS): R2070T | TRI Reagent

Supplemental Info.

• Tips & Tricks For RNA Isolation
• How to Extract RNA from TRIzol®
• How to Design Primers for PCR Experiments
• What is RNA-Seq?

FAQ

Q1: Is Direct-zol suitable for very small numbers of cells?

Yes, the Direct-zol MicroPrep (#R2060) is designed and capable of purifying RNA down to single cell inputs (picogram amounts). A sensitive quantification method is needed (e.g. Qubit, qPCR, etc.)

Q2: Is DNase I available for individual purchase?

All kit components are available for purchase separately.

Q3: How to store DNase-I following resuspension?

Lyophilized DNase I is stable at room temperature. Once resuspended, store frozen aliquots. Minimize freeze thaw cycles as much as possible. Freeze thaw will lower DNase activity.

Q4: Is the DNase-I treatment necessary?

If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.

Q5: Is the kit compatible with samples stored in DNA/RNA Shield?

Yes, bring samples homogenized and stored in DNA/RNA Shield to room temperature (20-30ºC). Add an equal volume (1:1) of TRIzol/TRI Reagent and mix well. Proceed with RNA Purification.

Q6: Is it possible to extract proteins with the Direct-zol RNA kits?

Yes, proteins can be acetone precipitated from the column flowthrough. Please see the Protein Purification appendix in the protocol.

Q7: Can samples be stored in TRIzol/TRI Reagent prior to processing?

Yes, samples in TRIzol/TRI Reagent or similar are stable overnight at room temperature and can be stored frozen (-80C). Be sure to lyse and homogenize the sample well prior to freezing. Bring the sample to room temperature prior to RNA Purification.

Q8: Is it possible to isolate DNA with the Direct-zol RNA kits?

Direct-zol DNA/RNA (R2080) kits can isolate DNA from TRIzol.

Q9: Is the RNA suitable for Next-Gen sequencing or other sensitive downstream applications?

Yes, the RNA is high quality (A260/A280 >1.8, A260/A230 >1.8) and suitable for any downstream application, including NGS, RT-PCR, hybridization, etc.

Q10: Which phenol-based reagents are compatible with Direct-zol?

The Direct-zol kits are compatible with TRI Reagent, TRIzol, Qiazol, RNAzol, TriPure, TriSure, etc., and any other acid-guanidinium phenol-based reagents.

Q11: What is the difference between the Direct-zol RNA and Quick-RNA kits?

Direct-zol is for samples stored/collected into TRIzol/similar reagents. Quick-RNA is for all other samples.

Q12: What is the difference between the Direct-zol RNA MiniPrep and the Direct-zol RNA MiniPrep Plus?

Both kits function the same, the only difference is the RNA binding capacity of the column provided with the kit.

Q13: I ran out of RNA Wash Buffer. Can I use something else?

Yes, use 80% ethanol as a substitute. RNA Wash Buffer is also sold separately.

Q14: Is this kit compatible with CTAB-based RNA extraction methods?

Yes, the Direct-zol RNA kits are compatible with CTAB-based RNA extraction methods for polysaccharide-rich and/or phenolics-rich samples (e.g., Pinus, Geranium plants). Please find detailed protocol here.

Q15: How to improve RNA yield?

• To ensure complete lysis, increase the volume of the lysis reagent (i.e., increase or titrate the volume of TRI Reagent/TRIzol or RNA Lysis Buffer). Lysate should be clear (not opaque or viscous). Pellet debris by centrifugation (if needed) and process the cleared supernatant.
• Prior to adding TRI Reagent/TRIzol or RNA Lysis Buffer, perform enzymatic treatment (Proteinase K; #D3001-2-20) and/or mechanical homogenization (bead beating with ZR BashingBead Lysis Tubes; #S6003) in DNA/RNA Shield (#R1100).

Reviews

"It took hardly any time, the protocol was so easy, and my RNA quality was SO much better. Honestly, this kit revolutionized my life at the bench."

A. Newhart

The Wistar Institute