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Plant/Fungi RNA Purification Kit FAQ

Product No. E4913

  1. Why am I experiencing poor RNA recovery?
  2. Why has my column become clogged?
  3. Why does my RNA not perform well in downstream applications?
  4. What volume of Lysis Solution should I use for extraction of RNA viroid from plants?
  5. How long can I store the sample after adding Lysis Solution before I complete the RNA purification?
  6. Can I use the Plant/Fungi RNA Purification Kit to isolate RNA from M. truncatula or A. thalina?
  7. I am obtaining a low yield of RNA. How can I increase the yield?
  8. My A260/230 ratio is low. How can I improve my RNA quality and increase the ratio?
  1. Why am I experiencing poor RNA recovery?

    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 solution was used - It is recommended that the Elution Buffer supplied with this kit be used for maximum RNA 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 50 mL of 95-100% ethanol is added to the supplied Wash Solution prior to use.

    Low RNA content in cells or tissues used - Different tissues and cells have different RNA contents, and thus the expected yield of RNA will vary greatly from these different sources. Please check literature to determine the expected RNA content of your starting material.

  2. Why has my column become 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 RNA content of the sample.

    Too much cell debris in the lysate supernatant - Ensure that most cell debris is removed in Step 1c of Lysate Preparation.

    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 my RNA not perform well in downstream applications?

    RNA was not washed 3 times with the provided Wash Solution - Traces of salt from the binding step may remain in the sample if the column is not washed 3 times with 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. What volume of Lysis Solution should I use for extraction of RNA viroid from plants?

    For 2 - 3 grams of leaf it is recommended that 600 µL of Lysis Solution should be used.

  5. How long can I store the sample after adding Lysis Solution before I complete the RNA purification?

    The sample can be stored for a few days at 4°C, a few weeks at -20°C and -70°C is recommended for storage of over a month.

  6. Can I use the Plant/Fungi RNA Purification Kit to isolate RNA from M. truncatula or A. thalina?

    Yes, the Plant/Fungi RNA Purification Kit can be used to isolate RNA from M. truncatula or A. thalina.

  7. I am obtaining a low yield of RNA. How can I increase the yield?

    Try using liquid nitrogen for the homogenization step to increase the yield.

  8. My A260/230 ratio is low. How can I improve my RNA quality and increase the ratio?

    An extra wash step can be performed to improve RNA quality.