Bio-Rad Chelex® 100 Molecular Biology Grade Resin - Manual (Page 5)

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2.2 DNA Preparation From Cultured Mammalian Cells
1
Pellet 200 µl of cell suspension from media in a microfuge tube. Spin
at 10,000–12,000 rpm for 1 minute and remove the supernatant.
2 Resuspend cells in 1 ml of 1x PBS and centrifuge for 1 minute at
10,000–12,000 rpm.
3
Resuspend cells in autoclaved water at 20–30 cells/µl.
4
Add 20 µl of this cell suspension to 200 µl of InstaGene matrix.
Incubate at 56 °C for 15–30 minutes.
NOTE:
InstaGene matrix should be mixed at moderate speed on a
magnetic stirrer to maintain the matrix in suspension. The pipet tip
used should have a large bore, such as a 1,000 µl pipet tip
(Bio-Rad’s catalog # 223-9378).
5
Vortex at high speed for 10 seconds. Place the tube in a 100 °C heat
block or boiling waterbath for 8 minutes.
6 Vortex at high speed for 10 seconds. Spin at 10,000–12,000 rpm for
2–3 minutes.
7
Use 20 µl of the resulting supernatant per 50 µl PCR reaction. Store
the remainder of the supernatant at -20 °C. Repeat step 6 when
reusing the InstaGene DNA preparation.
NOTE:
It is important to store the prepared sample at -20 °C.
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