What are the causes of not increasing amper in Non-denaturing PAGE?

There can be several reasons why the amperage does not increase during non-denaturing polyacrylamide gel electrophoresis (PAGE). Here are some possible causes to consider:

Incorrect voltage setting: Ensure that the voltage is set correctly on the power supply. The voltage should be sufficient to drive the migration of DNA fragments through the gel but not too high to cause excessive heating or damage to the gel. Check the voltage settings on the power supply and adjust if necessary.

Electrical circuit issues: Check the electrical connections in the gel apparatus, including the electrodes and cables. Make sure all connections are secure and not loose or damaged. Poor electrical connections can lead to inadequate current flow and a lack of increase in amperage.

Buffer conductivity: The conductivity of the buffer affects the electrical current during electrophoresis. Ensure that the 5X TBE buffer you prepared has the correct concentration of TBE salts to provide sufficient conductivity. If the buffer conductivity is too low, it can result in a lack of current increase. You can check the conductivity of the buffer using a conductivity meter and adjust it if needed.

Gel issues: Verify that the gel was properly prepared and cast. Ensure that the gel concentration is appropriate for the size range of your DNA fragments and that the gel is not too tight or too loose. An improperly prepared gel can affect the current flow and result in a lack of amperage increase.

Gel apparatus issues: Inspect the gel apparatus, including the chamber, electrode chambers, and buffer chambers. Ensure that there are no leaks or blockages that may impede the current flow. Make sure that the gel is properly positioned between the electrodes and that there are no air bubbles trapped in the gel or buffer.

Power supply limitations: Some power supplies have maximum current limits, and if the resistance of the gel is too high, the power supply may reach its maximum current output and not increase further. Check the specifications of your power supply to see if it has any current limitations that may be affecting the amperage increase.

By addressing these potential causes, you should be able to troubleshoot the issue of not increasing amperage during non-denaturing PAGE. It's important to follow proper gel electrophoresis protocols and optimize the conditions for your specific experiment to obtain the desired results.

These video playlists might be helpful to you:

https://www.youtube.com/playlist?list=PLzBiAPKi8vOPbE7Db9mKwloe34E8rXXEU

https://www.youtube.com/playlist?list=PLzBiAPKi8vONSqjcweFWHMRy13y3484bE

https://www.youtube.com/playlist?list=PLzBiAPKi8vOMheKwb0mu1dcZRaeGUFSlw

Does remaining RNA effects the cell cycle histogram in PI staining flow cytometry?

Validity or limits of einstein fields equations on torus?

MSc Student in Marine Biology, Population Genetics and Phylogeny

Protein-aptamer gel electrophoresis help, please??

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

How to regulate SSCP temperature in simple PAGE?

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

Why are my bands weird in this Agarose gel?

Which cell line should we use to perform biological dosimetry experiments?

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Calculating volume of DNA required from DNA concentration?

Has anyone used the Illumina ITS1 primer sets?