What is the meaning of having a DNA with good amount that is not visible on gel?

The most likely explanation is that you don't actually have plasmid at that concentration. I presume the gel was fine and your staining was good (otherwise that might be an answer).

If you the 50ng/ul comes from the A260 measurement, remember that degraded DNA or nucleotides can also give you an A260 peak. Or other contaminants can also absorb at A260 but that might not be the peak (hence we often use 260/280 ratios to test for purity). A lot of protein, or some phenol or some other contaminants can give you high A260 readings.

Mohammad Alzeyadi

Hi...

There could be a few reasons why you are not seeing any bands on your gel:

The concentration of your plasmid may be too low. If you are loading a large volume (e.g., 10 ul) of a sample with a low concentration (e.g., 50 ng/ul), it may be difficult to visualize the bands on the gel. You may need to concentrate the sample by either increasing the volume of plasmid DNA or decreasing the volume of sample loaded onto the gel.

The plasmid may have been damaged or degraded. If the plasmid DNA has been degraded or damaged, it may not produce a visible band on the gel. You may want to check the quality of your plasmid DNA using techniques such as agarose gel electrophoresis or a spectrophotometer.

There may be problems with the gel or the electrophoresis conditions. Make sure that the gel is properly prepared and that the electrophoresis conditions (e.g., voltage, time) are appropriate for your sample.

There may be issues with the loading dye. Make sure that the loading dye is prepared correctly and that it is mixed thoroughly with the sample before loading it onto the gel.

I hope this information is helpful!

Nikolay Klyashtornyy

If you have a DNA sample with a good amount (such as 50 ng/µL) but it is not visible on a gel, there are a few possible explanations:

Low sensitivity of the gel: Some gels, especially those with higher agarose concentrations or smaller pore sizes, may have lower sensitivity and may not detect low concentrations of DNA. Consider using a lower percentage agarose gel or a different gel system that is more suitable for visualizing lower concentrations of DNA.

Insufficient loading volume: Loading only 10 µL of a DNA sample, even if it has a high concentration, may not be sufficient to produce visible bands on the gel. Consider increasing the volume of your sample loaded onto the gel. However, be cautious not to overload the gel, as it can lead to smearing or distorted bands.

DNA degradation or damage: It is possible that the DNA in your sample has been degraded or damaged, which can result in no visible bands on the gel. Ensure that your DNA sample has been properly handled, stored, and prepared to maintain its integrity. Minimize exposure to UV light and excessive freeze-thaw cycles, and consider using freshly prepared DNA if possible.

Gel electrophoresis issues: Double-check the gel electrophoresis process, including the running buffer, voltage, and run time. Ensure that the gel is properly prepared, and there are no issues with the electrophoresis apparatus or power supply. Inadequate running conditions or equipment problems can result in poor separation and lack of DNA band visualization.

Loading buffer interference: The presence of contaminants or incompatible loading buffer components can interfere with the migration of DNA on the gel or affect the staining/visualization of the DNA bands. Ensure that your loading buffer is appropriate and free from any potential contaminants or inhibitors.

If you have ruled out technical issues and the DNA sample is not visible even after optimizing the gel conditions, it is possible that there may be an issue with the DNA itself, such as low quality or degradation. In such cases, it may be necessary to re-extract or purify the DNA sample to obtain better results.

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