Bio-Rad Chelex® 100 Molecular Biology Grade Resin - Manual (Page 6)

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2.3 DNA Preparation From Bacteria
The protocol described below is for the preparation of genomic DNA
or episomal DNA from bacteria.
1
Pick an isolated bacterial colony and resuspend it in 1 ml of
autoclaved water in a microfuge tube.
2 Centrifuge for 1 minute at 10,000–12,000 rpm. Remove the
supernatant.
3
Add 200 µl of InstaGene matrix to the pellet and 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).
4
Vortex at high speed for 10 seconds. Place the tube in a 100 °C heat
block or boiling waterbath for 8 minutes.
5 Vortex at high speed for 10 seconds. Spin at 10,000–12,000 rpm for
2–3 minutes.
6
Use 20 µl of the resulting supernatant per 50 µl PCR reaction. Store
the remainder of the supernatant at -20 °C. Repeat step 5 when
reusing the InstaGene DNA preparation.
NOTE:
It is important to store the prepared sample at -20 °C.
* The polymerase chain reaction (PCR) process is covered by U.S. patent numbers
4,683,195, 4,683,202, and 4,899,818 which are owned by Hoffman-La Roche, Inc. and
F. Hoffman- La Roche, Ltd. The purchase of this product does not convey a license to
use the process covered by these patents. The user of this product to perform PCR
must obtain a license from Hoffman-La Roche, Inc.
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