Jan 20, 2025

Public workspaceDNA Extraction with QIAGEN Mini Blood kit (modification for double swab)

This protocol is a draft, published without a DOI.
  • Paula Reyes1,
  • Alejandra Medina-Rivera2
  • 1UNAM;
  • 2Universidad Nacional Autonoma de Mexico
Paula Reyes
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Protocol Citation: Paula Reyes, Alejandra Medina-Rivera 2025. DNA Extraction with QIAGEN Mini Blood kit (modification for double swab). protocols.io https://protocols.io/view/dna-extraction-with-qiagen-mini-blood-kit-modifica-dpic5kaw
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: October 14, 2024
Last Modified: January 20, 2025
Protocol Integer ID: 109860
Keywords: dna extraction with qiagen mini blood kit, mini blood qiagen kit protocol, qiagen mini blood kit, dna extraction, modification for double swab, buccal swabs from the same individual, double swab, buccal swab, dna, extraction, procedure
Abstract
This procedure is modified from the Mini Blood QIAGEN kit Protocol and is intended to be used when two buccal swabs from the same individual are intended to be processed at the same time. The original protocol , can be consulted in https://qiagen.com/HB-0329.
Materials
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Troubleshooting
Before starting


Clean the area and pipettes using 70% Ethanol.
Take out 2 tubes of 2 mL for each pair of swabs to be processed. Label the tubes with the sample ID. Set the hotplate toTemperature85 °C

Extraction
1h 45m 30s
In a 2 mL tube, place Amount200 µL of PBS and Amount150 µL of ATL buffer, then add both swabs to the tube + vortex for Duration00:00:15 .

15s
Incubate atTemperature85 °C for Duration00:10:00 at Centrifigation1400 rpm and spin (30 seconds).

10m
Add Amount20 µL of proteinase K + vortex for Duration00:00:15 .

15s
Incubate at Temperature56 °C forDuration00:45:00 at Centrifigation1400 rpm and spinDuration00:00:30

45m 30s
Add Amount400 µL of AL buffer + vortex for Duration00:00:15

15s
Incubate atTemperature70 °C for 15 minutes atCentrifigation1400 rpm and spin Duration00:00:30

Precipitate with Amount400 µL of freezing cold absolute ethanol + vortex for Duration00:00:15 or with micropipette

15s
TransferAmount700 µL of the solution to the kit column, let it rest for Duration00:01:00 and centrifuge at Centrifigation8000 rpm for Duration00:02:00 .

3m
Transfer the column to a new collection tube. TransferAmount700 µL of the solution to the column, let it rest for Duration00:01:00 , and centrifuge at Centrifigation8000 rpm for Duration00:02:00 .

3m
Discard the flow-through and transfer the column to a clean collection tube.
Add Amount500 µL of AW2 buffer to the column, let it rest forDuration00:01:00 , and centrifuge at Centrifigation8000 rpm forDuration00:02:00 .

3m
Discard the flow-through and transfer the column to a clean collection tube.
Add Amount500 µL of AW2 buffer to the column, let it rest for Duration00:01:00 , and centrifuge at Centrifigation12000 rpm forDuration00:05:00 .

6m
Discard the flow-through and transfer the column to a clean 1.5-2 mL tube.
Add Amount65 µL of AE buffer to the column, incubate at Temperature30 °C for Duration00:15:00 .

15m
Centrifuge at Centrifigation6000 rpm for Duration00:02:00 .

2m
Add Amount65 µL of AE buffer to the column, incubate atTemperature30 °C for Duration00:15:00 .

15m
Centrifuge atCentrifigation6000 rpm for Duration00:02:00 .

2m
Store atTemperature-20 °C