Bioline ISOLATE II Plant DNA Kit, Руководство по эксплуатации продукта

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Plant DNA Kit
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Centrifugation speed was too
high Centrifuge at a maximum speed of 11,000 x g. Higher velocities
may cause DNA shearing.
LOW DNA YIELD
POSSIBLE CAUSE RECOMMENDED SOLUTION
Homogenization of plant material
insufficient For most species we recommend grinding with steel beads or
mortar and pestle (see section 7.2). For disruption of the cell
wall it is important to homogenize the plant material thoroughly
until the sample is ground to a fine powder. Instead of freezing in
liquid nitrogen, the sample can also be lyophilized and ground at
room temperature.
Suboptimal lysis buffer used Lysis efficiencies of Lysis Buffer PA1 (CTAB) and Lysis Buffer
PA2 (SDS) differ and depend on individual plant species (see
Table 1). Test both buffers side‑by‑side to determine optimal
detergent system for your plant sample.
Suboptimal lysis buffer volume
was used Cell lysis might be insufficient and too much DNA might get lost
during lysate clarification i.e. dry material absorbs too much lysis
buffer. Add additional lysis buffer and increase the volume of
Binding Buffer PB proportionally.
Extraction of DNA from plant
material during lysis was
insufficient
Increase incubation time in lysis buffer (up to overnight).
Reagents not applied correctly Ensure steps for reagent preparation are followed (see section
7.4).
Suboptimal elution The DNA can either be eluted in higher volumes or by repeating
the elution step up to three times (see section 7.4). Incubate
ISOLATE II Plant DNA Spin Column with Elution Buffer PG at
65°C for at least 5 minutes.
Check the pH of the elution buffer, which should be in the range
of pH 8.0– 8.5. To ensure correct pH, use supplied Elution Buffer
PG (5 mM Tris/HCl, pH 8.5).
SUBOPTIMAL PERFORMANCE OF EXTRACTED GENOMIC DNA IN ENZYMATIC REACTIONS
POSSIBLE CAUSE RECOMMENDED SOLUTION
Carry‑over of ethanol or salt Ensure the final two wash steps are performed with Wash
Buffer PAW2 and that the membrane was dried according to the
protocol.
Elution of DNA with buffers other
than Elution Buffer PG e.g. TE
Buffer We recommend elution with supplied Elution Buffer PG (5 mM
Tris/HCl, pH 8.5), as chemicals such as EDTA found in other
buffers can interfere with downstream applications.