From QIAamp DNA Mini & Blood Mini Handbook, Apendix B, p 50
Do not use more than 5 x 10^6 cells
Important points before starting
- Do not use more than 5 x 106 cells (with a normal set of chromosomes).
- All centrifugation steps are carried out at room temperature (15–25°C).
- Use carrier DNA if the sample contains <10,000 genome equivalents (see page 17).
Things to do before starting
- Heat a water bath or heating block to 56°C.
- Equilibrate Buffer AE or distilled water to room temperature (15–25°C) for elution.
- Ensure that Buffer AW1, Buffer AW2, and QIAGEN Protease have been prepared according to the instructions on page 16.
- If a precipitate has formed in Buffer AL, dissolve by incubating at 56°C.
Procedure
- Harvest cells
- Trypsinize and count cells
- Put no more than 5 million cells into a microfuge tube
- Centrifuge for 5 min at 300 x g.
- Remove the supernatant completely and discard, taking care not to disturb the cell pellet.
- Resuspend cell pellet in PBS to a final volume of 200μL.
- Add 20 μL QIAGEN Protease or proteinase K.
- (Continue with step 3 of “Protocol: DNA Purification from Blood or Body Fluids (Spin Protocol)”, page 27)
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