Q1: How to purify RNA from samples stored in RNAlater?
It is best to remove the RNAlater as it can interfere with the purification process: - For cells: Pellet via centrifugation at up to 5,000 x g and remove RNAlater (supernatant) - For tissue: Remove tissue using forceps, eliminate any excess reagent or crystals (e.g. PBS rinse) and proceed with addition of RNA Lysis Buffer. If RNAlater cannot be removed, add 1 volume of RNase-free water or PBS (1:1), then add 4 volumes DNA/RNA Lysis Buffer (4:1).
Q2: 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 1 volume of DNA/RNA Lysis Buffer (1:1) and mix well. Proceed with DNA Purification.
Q3: Can samples be stored in DNA/RNA Lysis Buffer prior to processing?
Yes, samples in DNA/RNA Lysis Buffer are stable overnight at room temperature and can be stored frozen (-80°C). Be sure to lyse and homogenize the sample well prior to freezing. Bring the sample to room temperature prior to RNA Purification.
Q4: What is the difference between the Quick-DNA/RNA Miniprep (D7001) and Quick DNA/RNA Miniprep Plus (D7003)?
Use the Quick-DNA/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).
Q5: 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.
Q6: Can I use other DNase I enzymes (or sets)? (e.g. Qiagen DNase I)?
The kit is optimized with Zymo’s DNase, however DNase from other manufacturers can be used. Follow the respective protocol for on-column DNase treatment. If the DNase does not have a protocol, proceed with in-tube DNase treatment post extraction, then purification using either “Reaction Clean-up procedure” or RCC-5.
Q7: 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.
Q8: Is Quick-DNA/RNA suitable for very small numbers of cells?
Yes, the Quick-DNA/RNA MicroPrep Plus (#D7005) 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.)
Q9: Is the kit compatible with blood samples stored in PAXgene Blood Tubes?
Yes, the kit is compatible with these anticoagulants.
Q10: Is the DNase-I treatment necessary?
If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.
Q11: Will the kit isolate small RNAs?
Yes, this kit will isolate small/micro RNA’s ≥ 17 nucleotides.
Q12: Is it possible to extract proteins with the Quick-DNA/RNA kit?
Yes, proteins can be Acetone Precipitated post RNA binding step. Please request supplementary protocol from Zymo Research Technical Support.
Q13: I ran out of DNA/RNA Wash Buffer. Can I use something else?
Yes. Use 80% ethanol as a substitute. DNA/RNA Wash Buffer is also sold separately.
Q14: Is the DNA and RNA suitable for Next-Gen Sequencing or other sensitive downstream applications?
Yes, both DNA and RNA is high quality (A260/A280 >1.8, A260/A230 >1.8) and suitable for any downstream application, including NGS, qPCR, etc.
