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@HB-2530-001 1113198 Pcard QIAamp DNA Mini 0418 WW

The document provides a quick-start protocol for extracting DNA from tissue samples using the QIAamp DNA Mini Kit. The 10 step protocol involves lysing the tissue, binding the DNA to a spin column, washing away contaminants, and eluting the purified DNA.

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29 views2 pages

@HB-2530-001 1113198 Pcard QIAamp DNA Mini 0418 WW

The document provides a quick-start protocol for extracting DNA from tissue samples using the QIAamp DNA Mini Kit. The 10 step protocol involves lysing the tissue, binding the DNA to a spin column, washing away contaminants, and eluting the purified DNA.

Uploaded by

rosita
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Quick-Start Protocol April 2018

QIAamp® DNA Mini Kit


The QIAamp DNA Mini Kit (cat. nos. 51304 and 51306) can be stored at room temperature
(15–25°C) for up to 12 months.

Further information

 QIAamp DNA Mini and Blood Mini Handbook: www.qiagen.com/HB-0329


 Safety Data Sheets: www.qiagen.com/safety
 Technical assistance: support.qiagen.com

Notes before starting

 Perform all centrifugation steps at room temperature (15–25°C).


 Use carrier DNA if the sample contains <10,000 genome equivalents. Refer to the
handbook for required equipment and procedure.
 If a precipitate has formed in Buffer ATL or Buffer AL, dissolve by incubating at 56°C.
 Add ethanol to Buffer AW1 and Buffer AW2 concentrates, as indicated on the bottle.
 Equilibrate samples to room temperature (15–25°C).
 Heat two water baths or heating blocks: one to 56°C for use in step 1 and one to 70°C
for use in step 3.
Refer to the handbook for protocols for swabs, dried blood spots, cultured cells, fixed
tissue, bacteria, yeast or other material.

Sample to Insight__
1. Cut tissue (≤25 mg) into small pieces and place in a 1.5 ml microcentrifuge tube. Add
180 µl Buffer ATL and 20 µl Proteinase K, mix by vortexing and incubate at 56°C until
completely lysed (1–3 h). Vortex occasionally during incubation.
2. Add 200 µl Buffer AL. Mix thoroughly by vortexing for 15 s.
3. Incubate at 70°C for 10 min. Briefly centrifuge the tube to remove drops from the lid.
4. Add 200 µl ethanol (96–100%). Vortex for 15 s. Briefly centrifuge the tube to remove
drops from the lid.
5. Pipet the mixture onto the QIAamp Mini spin column (in a 2 ml collection tube). Centrifuge
at 6000 x g (8000 rpm) for 1 min. Discard the flow-through and collection tube.
6. Place the QIAamp Mini spin column in a new 2 ml collection tube and add 500 µl
Buffer AW1. Centrifuge at 6000 x g (8000 rpm) for 1 min. Discard the flow-through
and collection tube.
7. Place the QIAamp Mini spin column in a new 2 ml collection tube and add 500 µl
Buffer AW2. Centrifuge at full speed (20,000 x g; 14,000 rpm) for 3 min. Discard the
flow-through and collection tube.
8. Recommended: Place the QIAamp Mini spin column in a new 2 ml collection tube (not
provided) and centrifuge at full speed for 1 min. This eliminates the chance of possible
Buffer AW2 carryover.
9. Place the QIAamp Mini spin column in a new 1.5 ml microcentrifuge tube (not provided),
add 200 µl Buffer AE or distilled water and incubate at room temperature for 1 min.
Centrifuge at 6000 x g (8000 rpm) for 1 min to elute the DNA.
10. Optional: Repeat step 10 for increased DNA yield with a further 200 µl Buffer AE or
distilled water.

Scan QR code for handbook.


For up-to-date licensing information and product-specific disclaimers, see the respective
QIAGEN kit handbook or user manual.
Trademarks: QIAGEN®, Sample to Insight®, QIAamp® (QIAGEN Group). 1113198 04/2018 HB-2530-001 © 2018 QIAGEN, all rights reserved.

Ordering www.qiagen.com/shop | Technical Support support.qiagen.com | Website www.qiagen.com

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