Bio-Rad Chelex® 100 Molecular Biology Grade Resin - Manual (Page 4)

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Section 2. Instruction for Use
2.1 DNA Preparation From Whole Blood
The protocol described below is for whole blood samples that are
frozen, refrigerated, or collected fresh.
1
Add 3–6 µl of whole blood to 1 ml of autoclaved nanopure water in a
1.5 ml microfuge tube. Mix by inverting the tube several times.
2 Incubate the tube at room temperature for 15–30 minutes.
3
Spin at 10,000–12,000 rpm for 2–3 minutes.
4
Carefully remove all but 20–30 µl of the supernatant. Do not disturb
the pellet.
5
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 TBR-78 tip, catalog # 223-9378).
6 Vortex at high speed for 10 seconds. Place the tube in a 100 °C heat
block or boiling waterbath for 8 minutes.
7
Vortex at high speed for 10 seconds. Spin at 10,000–12,000 rpm for
2–3 minutes.
8 Use 20 µl of the resulting supernatant per 50 µl PCR reaction. Store
the remainder of the supernatant at -20 °C. Repeat step 7 when
reusing the InstaGene DNA preparation.
NOTE: It is important to store the prepared sample at -20 °C.
NOTE:
For more efficient DNA preparation from whole blood, the
AquaPure T M Genomic DNA Blood Kit (catalog # 732-6345) is
recommended.
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