AmplifyRP FAQ

What advantages does AmplifyRP offer compared to conventional or real-time PCR?

Several.  Although PCR is a highly sensitive and specific type of DNA testing, it does require some skill and is typically restricted to laboratory environments.  In most PCR assays it is first required that suspect samples undergo a DNA purification or "clean-up" step.  This step requires the use of expensive purification kits and sometimes toxic chemicals.  PCR also requires an important step called thermal cycling where reactions are repeatedly subjected to temperature fluctuations that facilitate the actual PCR process.  To summarize, PCR requires expensive equipment, skilled technicians, and several hours to complete.

AmplifyRP provides a DNA testing platform that rivals PCR in terms of sensitivity and specificity, but it eliminates the need for expensive equipment and can be completed by users of all skill levels.... all in about 30 minutes.

  • • The total cost for equipment is as low as $300 depending on the format
  • • Crude extracts may be used for analysis
  • • Total assay time is between 25 and 40 minutes, including sample preparation
  • • Format is user-friendly and can be field portable

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Do I need special training to use AmplifyRP?

No.  AmplifyRP was designed with multiple types of end users in mind.  All the reagents and disposable equipment necessary to run an AmplifyRP test comes included in a kit.  Any equipment required can be purchased upfront in a starter pack.

AmplifyRP kits were also designed to limit the number of steps in the protocol.  All the amplification reagents are lyophilized in one-time use tubes.  The only thing the end user needs to do is rehydrate the reagents and insert their sample.  Once that is complete, amplification and detection can be completed in 15 to 30 minutes depending on the format.

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What part of my ground sample should I use to test?

With your sterile loop or pipette, please take your extract from the fully mixed sample solution. Do not add large pieces of tissue to the reaction.

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What happens if I add more than prescibed amount of extract to the reaction?

The amount of sample extract that you add to a reaction is dependent on the test you are running.  Refer the assay user guide. Small changes to volume added (1ul or less) do not affect test performance.

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Can I use an extraction buffer other than the recommended buffer in my test?

Only the extraction buffer supplied with the kit are validated with the test. Grinding tissue in any other buffer may lead to false negative results.

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What happens if I incubate the reaction at a high temperature?

If you incubate your reaction at a temperature significantly higher than 39°C (42°C or greater), the enzymes in the pellet will become inactive increasing the chance of a false negative result.

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What happens if I incubate the reaction at a low temperature?

If you incubate your reaction at a temperature lower than 39°C, the reaction will occur more slowly. Since a lower temperature incubation requires a longer incubation time, adding the reaction to the detection chamber too soon can lead to a false negative result.

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What happens is the heat supply has been interrupted?

If your heat supply has been interrupted during the reaction or if you’ve mixed your extract with the reaction pellet without immediately heating the reaction, the test may still be valid. After you have noticed that the heat supply has been interrupted or the heat block was not turned on, place your reaction back in your heat source for an additional 15 minutes and proceed with the instructions as recommended.

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Do you have a protocol for insect or other tissues?

If an extraction protocol for your tissue type is not specifically described in your user guide, please contact techsupport@agdia.com or 1-800-622-4342 for more information.

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Can I retrieve the strip from the detection chamber for my own records?

Once you have closed the detection chamber, do not attempt to re-open the device. Opening or mishandling the chamber from a completed reaction could lead to contamination of the work area and false positive results. If you would like to keep a record of the results, Agdia recommends keeping a photographic record.

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What should I do if my reaction tube lid pops open before I secure it in the chamber?

If a tube from a completed reaction opens before it is secured in the detection chamber, please discard the test and clean the area. If you suspect you have exposed your work area to the contents of a completed reaction, wash your hands and wipe nearby surfaces with a bleach solution before performing your next test.

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How do I dispose of the completed test reaction?

The detection chamber and associated materials can be disposed as general waste. Do not disassemble the detection chamber.

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Does this test include an internal reporter (housekeeping) control like some PCR reactions?

The control line in an Acceler8 assay is a measure of the performance of the lateral flow strip, not the nucleic acid extraction. Internal reporters or housekeeping primers are used in PCR reactions to detect the presence of host (plant) nucleic acid following nucleic acid purification. AmplifyRP technology utilizes crude plant extracts and does not require any purification technique that could lead to the loss of host DNA or RNA, so internal reporters are not used.

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What should I do if the product appears damaged?

Contact Agdia immediately if you notice any defect to the packaging or kit components or missing item.

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Who can I contact if I have questions about the test performance?

Please contact Agdia Technical Support at techsupport@agdia.com or 1-800-622-4342.

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