Direct-zol-96 RNA Kits
R2054 / R2055 / R2056 / R2057
Direct-zol-96 RNA Kits
- 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.
The Direct-zol-96 RNA Kits are RNA purification kits that provide a streamlined method for the purification of up to 10 µg (per well) of high-quality RNA directly from samples in TRI Reagent®. 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.) using this product. The extraction method inactivates viruses and other infectious agents.
The procedure is easy: simply apply a sample in TRI Reagent® to the Zymo-Spin I-96 Plate, then bind, wash, and elute the RNA. No phase separation, precipitation, or post-purification steps are necessary. 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 30 minutes (per 2 plates).
|Compatibility||TRIzol®, RNAzol®, QIAzol®, TriPure™, TriSure™ and all other acid-guanidinium-phenol based solutions can be used in place of TRI Reagent®.|
|Equipment||Microplate centrifuge, vortex|
|Sample Inactivation||TRI Reagent® (provided with R2055, R2057) 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||10 µg RNA (binding capacity), ≥10 µl (elution volume)|
Q1: Is DNase I available for individual purchase?
All kit components are available for purchase separately.
Q2: 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.
Q3: Is the DNase-I treatment necessary?
If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.
Q4: 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 3 volume of TRIzol/TRI Reagent and mix well. Proceed with RNA Purification.
Q5: 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.)
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: What is the average RNA yield? (chart)
|Input||Average RNA Yield|
|Cells||10 µg (per 106 cells)|
|High Yield Tissue (mouse)||≥ 30 µg (per 10 mg)|
|Low Yield Tissue (mouse)||≤ 30 µg (per 10 mg)|
|Brain, Heart||5-15 µg|
|Whole Blood||(per 1 ml)|
Q15: How to improve purity, RIN scores and eliminate contamination (i.e., A260/230, A260/280 ratios, DNA, phenol, protein, salt, etc.)?
Purity, RIN and/or any type of contamination can result from initial sample preparation (i.e., inefficient lysis of the sample). To improve, increase the volume of the lysis reagent (e.g., TRI Reagent/TRIzol or RNA Lysis Buffer).
Q16: 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).
“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.”
- Adina B. (University of Guelph)
“This kit is amazing, 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!“
- R.K. CSU
“Direct-zol is the most excellent kit for RNA isolation that I ever used in the past 20 years.”
- H.Z. (Harvard Medical School)
|E1010-1-16||DNA Digestion Buffer||16 mL|
|R2050-1-200||TRI Reagent||200 ml|
|E1010-1-4||DNA Digestion Buffer||4 mL|
|C2003||Elution Plate||2 Plates|
|C2002||Collection Plate||2 Plates|
|C2007-2||96-Well Plate Cover Foil||2 Foils|
|C2007-4||96-Well Plate Cover Foil||4 Foils|
|C2004||Zymo-Spin I-96 Plate||2 Plates|
|W1001-30||DNase/RNase-Free Water||30 ml|
|W1001-10||DNase/RNase-Free Water||10 ml|
|R2050-2-160||Direct-zol RNA PreWash (Concentrate)||160 ml|
|R2054||Direct-zol-96 RNA||2 x 96 preps|
|R2056||Direct-zol-96 RNA||4 x 96 preps|
|R1003-3-48||RNA Wash Buffer||48 ml|
|E1010||DNase I Set||250 U|