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Plant/Fungi DNA Isolation Kit FAQ

Product No. E5038

  1. Why is my yield of genomic DNA low?
    Column has become clogged - Do not exceed the recommended amounts of starting materials. The amount of starting material may need to be decreased if the column shows clogging below the recommended levels. See also “Clogged Column” below.

    An alternative elution buffer was used - It is recommended that the Elution Buffer supplied with this kit be used for maximum DNA recovery.

    Lysis Additive was not added to the lysate - Ensure that the provided Lysis Additive is added to the lysate and that the incubation at 65°C for 10 minutes is performed to maximize DNA recovery.

    Ethanol was not added to the lysate - Ensure that the appropriate amount of ethanol is added to the lysate before binding to the column.

    Ethanol was not added to the Wash Solution - Ensure that 70 mL of 95 - 100% ethanol is added to the supplied Wash Solution prior to use.

  2. Why is my spin column becoming clogged?
    Maximum number of cells or amount of tissue exceeds kit specifications - The optimal input is 50 mg of plant tissue or filamentous fungi, or 5 x 106 plant cells. However, for some species, up to 100 mg of tissue may be processed depending on the DNA content of the sample.

    Too much cell debris in the lysate supernatant - Ensure that most cell debris is removed in Steps 1e and 1f.

    Centrifuge temperature too low - Ensure that the centrifuge remains at room temperature throughout the procedure. Temperatures below 20°C may cause precipitates to form that can cause the columns to clog.

  3. Why does the DNA not perform well in downstream applications?
    PCR reaction condition is needed to be optimized - Take steps to optimize the PCR conditions being used, including varying the amount of DNA template, changing the source of Taq polymerase, looking into the primer design and adjusting the annealing condition.

    Binding Solution was not added to the lysate - Ensure that the Binding Solution is added to the lysate and that it is incubated on ice for 5 minutes prior to spinning down the lysate.

    Lysis Additive was not added to the lysate - Ensure that the provided Lysis Additive is added to the lysate and that the incubation at 65°C for 10 minutes is performed to maximize DNA recovery.

    DNA was not washed twice with the provided Wash Solution - Traces of salt from the binding step may remain in the sample if the column is not washed twice with the Wash Solution. Salt may interfere with downstream applications, and thus must be washed from the column.'

    Ethanol carryover - Ensure that the dry spin under the Column Wash procedure is performed, in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.

  4. I am using the kit to isolate DNA from pomegranate, which is known to be a difficult sample. What can I do to improve the yield and purity of my DNA? 
    We recommend using a smaller amount of input, between 25 and 30 mg. Also, the volume of Wash Solution can be increased from 500 up to 700 µL.

  5. Can I isolate DNA from wheat flour using this kit? 
    Yes, DNA can be isolated from as little as 5 mg of wheat flour using this kit.

  6. Can I isolate DNA from blueberries and strawberries for genotyping analysis using this kit? Yes, DNA can be isolated from both blueberries and strawberries using this kit.