Mar 18, 2026

Public workspaceZymoBIOMICS™ DNA/RNA Miniprep Kit

This protocol is a draft, published without a DOI.
  • Kristine Wylie1,
  • Jane Schrimpf1,
  • Madison Eschbach1,
  • Hunter Olson1
  • 1Washington University
Kristine Wylie
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Protocol Citation: Kristine Wylie, Jane Schrimpf, Madison Eschbach, Hunter Olson 2026. ZymoBIOMICS™ DNA/RNA Miniprep Kit. protocols.io https://protocols.io/view/zymobiomics-dna-rna-miniprep-kit-jv7ycn9px
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 16, 2026
Last Modified: March 18, 2026
Protocol Integer ID: 313304
Keywords: rna miniprep kit, rna miniprep kit protocol, zymobiomics dna, micro rna, free total rna
Funders Acknowledgements:
NIH NCCIH
Grant ID: U01 AT012970
Abstract
Protocol was adapted from the ZymoBIOMICS DNA/RNA Miniprep Kit – For 50 preps: Purification of high quality, inhibitor-free total RNA (including small/micro RNAs). Catalog number: R2002
Materials
  • Microcentrifuge
  • Pipettor (50–200 µL; 100–1000 µL)
  • Vortex-Genie 2 Vortex
  • Vortex Adapter for 24 (1.5–2.0 mL) tubes (cat. no. 13000-V1-24)
Troubleshooting
Before start
  • Store unopened kit at room temperature.
  • Wear a lab coat, face mask, and nitrile gloves while extracting DNA and RNA from swabs.
  • All centrifugation performed @ 16,000 x g for 30 seconds unless otherwise specified.
  • All steps done at room temperature unless specified.
Reagent and Tube Preparation
Before using a kit, add 96 mL 100% ethanol to the 24 mL RNA Wash Buffer.
Before using a kit, add 275 µL DNase/RNase-Free Water per vial to reconstitute the lyophilized DNase I @ 1 U/µL. Mix gently by inversion.
Add 400 µL of DNA/RNA Lysis Buffer to a 1.7 mL RNase-free tube (not provided).
Place a Spin-Away Filter (yellow) in a collection tube and label the top of column and the side of the collection tube.
Place a Zymo-Spin IIICG column (green) in a collection tube.
Sample Preparation
Write the sort # of the sample on top of each swab tube to mark it as extracted.
Thaw the ZR BashingBead Lysis tubes which contain samples in DNA/RNA Shield at RT.
Secure the ZR BashingBead Lysis tube horizontally using the Vortex Adapter tube holder for the vortex. Vortex at maximum speed for 15 minutes.
Centrifuge for 30 sec.
Transfer 400 µL of supernatant to the tube containing DNA/RNA Lysis Buffer and mix. Return the ZR BashingBead Lysis tube containing the remains of the sample to its original spot in the -80 C freezer.
DNA/RNA Purification
Transfer 800 µL mixture into the Spin-Away Filter (yellow) and centrifuge. RNA is in flow-through, and DNA is bound to column.
Store flow-through containing RNA at RT and proceed immediately with DNA purification from column.
DNA Purification
Transfer the yellow Spin-Away Filter into a new collection tube.
Add 400 µL DNA/RNA Prep Buffer to column and centrifuge. Discard flow-through.
Add 700 µL DNA/RNA Wash Buffer to column and centrifuge. Discard flow-through.
Add 400 µL DNA/RNA Wash Buffer to column and centrifuge for 2 min. Transfer the column carefully into a 1.7 mL RNase-free tube (not provided).
Add 100 µL DNase/RNase- Free Water directly to the column, incubate for 5 min. and centrifuge. Discard column and store eluted DNA on ice.
RNA Purification
Add an equal volume (770 µL) of ethanol (95-100%) to flow-through and mix.
Transfer the mixture into a Zymo-Spin IIICG column (green) in a collection tube and centrifuge. Discard flow-through. Repeat until entire sample has been put over column.
Add 400 µL DNA/RNA Wash Buffer to column and centrifuge. Discard flow-through.
For each sample to be treated prepare DNase I Reaction Master Mix in microfuge tube:


Note: Mix by pipetting gently. Try to avoid making any bubbles.
Add 80 µL DNase I Reaction Mix directly onto the column matrix and incubate at room temperature for 15 minutes.
Add 400 µL DNA/RNA Prep Buffer to column and centrifuge. Discard flow-through.
Add 700 µL DNA/RNA Wash Buffer and centrifuge. Discard flow-through.
Add 400 µL DNA/RNA Wash Buffer and centrifuge for 2 min. Discard the flow-through. Carefully transfer the column into a 1.7 mL RNase-free tube (not provided).
Add 100 µL DNase/RNase- Free Water directly to the column, incubate for 5 min. and centrifuge. Store eluted RNA on ice.
Filtration of Purified DNA/RNA
For each sample, place a Zymo-Spin III-HRC Filter in a Collection Tube and add 600 µL ZymoBIOMICS HRC Prep Solution. Centrifuge at 8,000 x g for 3 minutes.
Transfer the eluted DNA and RNA into a prepared Zymo-Spin III-HRC Filter in a 1.7 mL stock elution tube with label and centrifuge at exactly 16,000 x g for 3 minutes.
Place eluted DNA and RNA on ice.
Pipet 50 µL of DNA or RNA into the library aliquot tube.
Quality Control
Using sample in stock tube, determine DNA and RNA concentration with dsDNA HS and RNA HS Qubit Assay kits.
Store DNA and RNA stock tubes and library aliquot tubes at -80°C.