The Quick-RNA Miniprep Plus Kit is one of the most innovative RNA isolation kits available, designed for the easy, reliable, and rapid isolation of DNA-free RNA from cells, all tissue types, whole blood, and biological fluids. The provided DNA/RNA Shield stabilizes samples, allowing them to be stored without the need for immediate freezing or processing for up to one month. Furthermore, DNA/RNA Shield™ inactivates RNases as well as microbial pathogens (viruses, bacteria, etc.). The procedure combines a unique buffer system with Zymo-Spin Column technology to yield high-quality total RNA (including small RNAs 17-200 nt). Simply add DNA/RNA Shield and Proteinase K to extract total RNA from any tissue, then purify the RNA using the Zymo-Spin Column. The result is highly concentrated, DNA-free RNA that is suitable for RT-PCR, hybridization, sequencing, etc. In addition, the kit can be used for the enrichment of small and large RNAs in two separate fractions.
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|Equipment||Microcentrifuge, vortex, heat block/bath (55ºC)|
|Purity||RNA is ready for Next-Gen sequencing, RTPCR, microarray, hybridization, etc. A260/A280, A260/A230: >1.8.|
|Sample Source||Any cells (animal, blood cells etc.), all tissues (tough-to-lyse, FFPE, etc.), blood, biological fluids, and enzymatic reactions (e.g., DNase I), samples in DNA/RNA Shield™, TRIzol® or RNAlater™.|
|Size Range||Total RNA ≥ 17 nt|
|Yield||spin column, 100 µg RNA (binding capacity), ≥50 µl (elution volume)|
Q1: What is the difference between the Quick-RNA Miniprep and the Quick-RNA Miniprep Plus?
Use the Quick-RNA Miniprep for cells and soft tissues. The “Plus” kit accommodates all sample types (cells, tissue, blood) and comes with DNA/RNA Shield (sample collection, transport, storage at ambient temperature).
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 DNase I available for individual purchase?
Yes, the catalog number for the DNase I set (DNase and DNA Digestion Buffer) that we offer is E1010.
Q5: 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.
Q6: Will the kit isolate small RNAs?
Yes, this kit will isolate small/micro RNA’s ≥ 17 nucleotides.
Q7: Can samples be stored in RNA Lysis Buffer prior to processing?
Yes, samples in RNA Lysis Buffer 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: 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.
Q9: Is it possible to extract proteins with the Quick-RNA kit?
Yes, proteins can be acetone precipitated from the column flowthrough. Please request supplementary protocol from Zymo Research Technical Support.
Q10: 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)|
Q11: 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).
Q12: 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).