Direct-zol-96 MagBead RNA
R2103 / R2102 / R2100 / R2101
Direct-zol-96 MagBead RNA
- 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 MagBead RNA kit provides a high-throughput (96-well), magnetic bead-based isolation of high-quality RNA directly from samples in TRI Reagent® and similar. The extraction method inactivates viruses and other infectious agents. Total RNA including small/microRNAs (≥17 nt) are effectively isolated from a variety of sample sources (cells, tissues, biological liquids, etc.). The procedure is easy: simply add ethanol and MagBinding Beads to a sample in TRI Reagent®, 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 Next-Gen sequencing, RT-PCR, transcription profiling, hybridization, etc.
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|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, magstand|
|Sample Inactivation||TRI Reagent® (provided with R2101, R2103, and R2105) 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), ≥ 30 µl (elution volume)|
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: 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).
“Before I discovered this kit, I was isolating RNA the old school way with chloroform and it would take half the day to finish the protocol. The Direct-zol RNA Miniprep kit is AWESOME! 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)
“Simple protocol and yielded good quality of RNA. Only one kit working for all type of tissue, cell and especially biological fluids.”
- Mohan K. (University of Illinois, Chicago)
“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? Just one column and two buffers, I love it.”
- Arjan V. (Indiana University)
|D4100-2-3||MagBinding Beads||3 ml||$72.60|
|D4100-2-12||MagBinding Beads||12 ml||$137.50|
|W1001-30||DNase/RNase-Free Water||30 ml||$24.20|
|R1060-2-100||RNA Prep Buffer||100 ml||$134.20|
|R1060-2-25||RNA Prep Buffer||25 ml||$44.00|
|R2130-1-120||MagBead DNA/RNA Wash 1||120 ml||$217.80|
|R2130-1-30||MagBead DNA/RNA Wash 1||30 ml||$69.30|
|R2130-2-20||MagBead DNA/RNA Wash 2||20 ml||$59.40|
|R2130-2-80||MagBead DNA/RNA Wash 2||80 ml||$188.10|
|C2002||Collection Plate||2 Plates||$24.20|
|C2003||Elution Plate||2 Plates||$20.90|
|C2007-8||96-Well Plate Cover Foil||8 Foils||$18.00|
|R2050-1-200||TRI Reagent||200 ml||$248.60|
|E1010-1-4||DNA Digestion Buffer||4 mL||$16.50|
|E1010||DNase I Set||250 U||$61.60|