Mar 17, 2026

Public workspaceQIAamp DNA Blood Mini Kit

This protocol is a draft, published without a DOI.
  • Kristine Wylie1,
  • Jane Schrimpf1,
  • Madison Eschbach1,
  • Hunter Olson1
  • 1Washington University
  • HVP Pregnancy Postpartum
Hunter Olson
Icon indicating open access to content
QR code linking to this content
Protocol Citation: Kristine Wylie, Jane Schrimpf, Madison Eschbach, Hunter Olson 2026. QIAamp DNA Blood Mini Kit. protocols.io https://protocols.io/view/qiaamp-dna-blood-mini-kit-jwadcpaa7
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 17, 2026
Protocol Integer ID: 313381
Keywords: qiaamp dna blood mini kit protocol, qiaamp dna blood mini kit, qiagen catalog number
Funders Acknowledgements:
NIH NCCIH
Grant ID: U01 AT012970
Abstract
Protocol was adapted from the QIAamp DNA Blood Mini Kit - Qiagen catalog number: 51104
Guidelines
  • QIAamp Mini spin columns and buffers can be stored dry at room temperature (15–25°C) for up to 1 year without showing any reduction in performance. 
  • Lyophilized QIAGEN Protease can be stored at room temperature for up to 12 months without any decrease in performance. For storage longer than 12 months or if ambient temperatures constantly exceed 25°C, QIAGEN Protease should be stored dry at 2–8°C.
  • QIAGEN Protease reconstituted in Buffer AVE, Protease Solvent and Protease Suspension buffer is stable for 12 months when stored at 2–8°C. Keeping the QIAGEN Protease stock solution at room temperature for prolonged periods of time should be avoided.
Materials
  • Microcentrifuge
  • Pipettor (50–200 µL; 100–1000 µL)
  • Water bath or heating block heated to 56°C.
Troubleshooting
Before start
  • Wear a lab coat, face mask, eye protection and nitrile gloves while extracting DNA from blood.
  • All centrifugation performed @ 6000 x g for 1 min. unless otherwise specified.
  • All steps done at room temperature unless specified.
Reagent and Tube Preparation
Add the appropriate amounts of ethanol (96-100%) to Buffer AW1 and Buffer AW2 concentrates.
Pipet 1.2 mL protease solvent into the vial containing lyophilized QIAGEN Protease, as indicated on the label, and store at 4°C (top shelf of door of refrigerator).
Sample Preparation
Equilibrate samples to room temperature.
Heat a water bath or heating block to 56°C.
If a precipitate has formed in Buffer AL, dissolving by incubating at 56°C.
Lysate Preparation
Pipet 20 µL of QIAGEN Protease into the bottom of a 1.7 mL microfuge tube.
Add 200 µL whole blood to the tube.
Add 200 µL Buffer AL to the sample. Mix by pulse-vortexing for 15 sec.
Note: Buffer AL contains guanidine hydrochloride so DO NOT combine with bleach.
Incubate at 56°C for 10 min.
Briefly centrifuge the tube to remove drops from the lid.
Add 200 µL ethanol to the sample and mix again by pulse-vortexing for 15 sec.
Briefly centrifuge the tube to remove drops from the lid.
Binding DNA to Column
Carefully apply the sample to QIAamp Mini spin column in a 2 mL collection tube without wetting the rim.
Close the cap and centrifuge at 6000 x g for 1 min. Place the column in a new collection tube and discard tube containing the flowthrough.
Note: Column may be centrifuged at higher speed if the entire sample has not passed through the column.
Add 500 µL Buffer AW1 without wetting the rim. Centrifuge at 6000 x g for 1 min. Place the column in a clean 2 mL collection tube and discard the collection tube containing the flowthrough.
Add 500 µL Buffer AW2 without wetting the rim. Centrifuge at full speed for 3 min.
Place the column in a new collection tube (not provided) and discard tube containing the flowthrough. Centrifuge at full speed for 1 min.
DNA Elution
Place the column in a labeled 1.7 mL microfuge tube (not provided) and add 200 µL Buffer AE or distilled water.
Incubate at RT for 5 min.
Centrifuge column at 6000 x g for 1 min.
Pipet eluted DNA into the QC tube (5 µL/tube), and the library aliquot tube (50 µL/tube).
Quality Control
Using sample in QC tube, determine DNA concentration with dsDNA HS Qubit Assay kit.
Store all tubes at -80°C.