FAQ

Frequently asked questions

This is frequently asked questions(FAQ) Inquiries for all Seegene products

Real-time PCR

When the Internal Control Signal is invisible

First, please check the selection of the correct fluorophore for data analysis and check whether the testing was conducted in accordance with the protocol guidelines. If there are no abnormalities, it may be a problem of PCR inhibition from the PCR inhibitor or a problem in extraction.
It is recommended that you conduct PCR again by diluting the 10-to-100 times extracted nucleic acid for any PCR inhibitor problem, test again with another aliquot of the same sample, or retest after collecting a new sample. In addition, please use a new kit when an abnormality is detected after checking the storage conditions and shelf life of the reagents.

When the signal is not observed in Negative Control or there is a false positive result

There is the possibility of cross contamination during the extraction process or PCR process. After sterilizing the testing area and the instrument with sodium hypochlorite and ethanol, it is recommended that you start with nucleic acid extraction again. Please re-test using new supplies and reagents if necessary.
To prevent contamination, please make sure to use the filter tip when testing, and regularly clean the testing area. Real-time PCR in the use of CFX96 or AB7500 instruments is conducted when the detection of fluorophore passes the cap of a tube. For marking using a pen, please make sure to use the surface of a real-time PCR tube.

When the signal is not observed in Positive Control or there is a false negative result

First, please check the selection of the correct fluorophore for data analysis and whether the testing was conducted in accordance with the reagent shelf life and within the protocol guidelines. In addition, please use a new kit when an abnormality is detected after checking the storage conditions and the shelf life of any reagents.

Conventional PCR testing

When the Internal Control Band is not visible

First, when there is no abnormality after checking whether the testing was conducted in accordance with the guidelines, inhibition is possible due to the PCR inhibitor. Please retry PCR after diluting the specimen or template DNA 10-to-100 times. In addition, use a new kit when an abnormality is detected after checking the storage conditions and shelf life of the reagents.

When PCR band isn't visible on Agarose gel

You can't confirm the band if Et-Br staining wasn't properly done or it's been overexposured to UV light . If Et-Br staining wasn't properly done, shake 1/10,000 dilution 10mg/ml Et-Br for 30 mins in 1100 ± 100 rpm shaker.

When the Band is not observed in Negative Control or there is a false positive result

Cross contamination is possible during the extraction process or PCR process. After sterilizing the testing area and the instruments with sodium hypochlorite and ethanol, it is recommended to start with nucleic acid extraction again. Please re-test using new supplies and reagents if necessary. To prevent contamination, please make sure to use the filter tip when testing, and regularly clean the testing area.

When the Band is not observed in Positive Control or there is a false negative result

Please check whether the testing was conducted in accordance with the shelf life of a reagent and within the protocol guidelines. (Sensitivity can be inhibited with abruptly decreasing enzyme efficacy when stored in a Premix form in advance. Please make sure to conduct the testing in accordance with the testing guidelines.)

When the specimen or nucleic acid is stored at room temperature for a long time

Low titer specimens or nucleic acids may show degradation after only a day or so, which may have an impact on the result. Never store specimens or nucleic acids at room temperature, but if so inevitably, please conduct the testing by re-extraction.

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