Bioline ISOLATE II Plant DNA Kit, Product Manual

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Product Manual www.bioline.com/isolate
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3 Filter crude lysate
Place an ISOLATE II Filter (violet) into a new Collection Tube (2 mL) and load lysate
onto column. Centrifuge for 2 min at 11,000 x g. Collect the clear flow‑through and
discard the ISOLATE II Filter.
Repeat the centrifugation step if not all liquid has passed through the filter.
If a pellet is visible in the flow‑through, transfer the clear supernatant without
disturbing the pellet to a new 1.5 mL microcentrifuge tube (not supplied).
4
Adjust DNA binding conditions
Add 450 μL Binding Buffer PB. Mix thoroughly by pipetting up and down 5 times or
by vortexing.
5 Bind DNA
Place an ISOLATE II Plant DNA Spin Column (green) into a new Collection Tube (2
mL) and load sample (max. of 700 μL).
Centrifuge for 1 min at 11,000 x g and discard the flow‑through.
The maximum loading capacity of the ISOLATE II Plant DNA Spin Column is 700 μL.
For higher volumes repeat the loading and centrifugation steps.
6 Wash and dry silica membrane
• Add 400 μL Wash Buffer PAW1 to the ISOLATE II Plant DNA Spin Column.
Centrifuge for 1 min at 11,000 x g and discard flow‑through.
Note: Although washing with Wash Buffer PAW1 increases purity, it can in certain cases reduce
the final yield slightly.
• Add 700 μL Wash Buffer PAW2 to the ISOLATE II Plant DNA Spin Column.
Centrifuge for 1 min at 11,000 x g and discard flow‑through.
• Add another 200 μL Wash Buffer PAW2 to the ISOLATE II Plant DNA Spin
Column. Centrifuge for 2 min at 11,000 x g in order to remove wash buffer and
to dry the silica membrane completely.
7
Elute DNA
Place the ISOLATE II Plant DNA Spin Column into a new 1.5 mL microcentrifuge tube
(not supplied).
Pipette 50 μL Elution Buffer PG (65°C) onto the membrane. Incubate the ISOLATE II
Plant DNA Spin Column for 5 min at 65°C. Centrifuge for 1 min at 11,000 x g to elute
the DNA.
Repeat this step with another 50 μL Elution Buffer PG (65°C) and elute into the same
tube.
Note: To achieve maximum yield or higher concentrations refer to section 7.4 for alternative
elution procedures.