I am isolating DNA from Streptomyces species. The problem I am facing is when i am quantifying the DNA by using nanodrop the purity shows 1.8 ratio (260/280) and the concentration was also good like 250ng. But when i run gel for the same sample. It doesn't show any band on Gel.
Nikolay Klyashtornyy
Insufficient DNA: Even if your Nanodrop readings indicate that you have a good concentration of DNA, it is possible that the amount of DNA you have is not sufficient for visualization on a gel. You can try running a higher amount of DNA on the gel to see if this helps. Poor quality DNA: While the 260/280 ratio can be used as an indicator of DNA purity, it is not always reliable. It is possible that the DNA you have isolated is of poor quality and may not be suitable for gel electrophoresis. You may want to consider performing additional quality checks, such as checking the DNA integrity on an agarose gel, before proceeding with downstream applications. Gel electrophoresis issues: There could be issues with your gel electrophoresis setup. Make sure that your gel is prepared correctly and that your running buffer is fresh. You may also want to check that your power supply is working properly and that the electrodes are connected properly. Ethidium Bromide issues: Ethidium bromide is the most common DNA stain used for visualizing DNA on a gel. Make sure that you have added the appropriate amount of ethidium bromide to your gel and that it has been properly incorporated. You can also try using alternative DNA stains, such as SYBR Safe, to see if this helps with visualization. Other factors: Other factors that could contribute to not seeing any bands on the gel include degradation of DNA during isolation, DNA damage during extraction, or issues with the PCR amplification (if you are using PCR to generate DNA fragments).
There are a few possible reasons why you might not be seeing any bands on the gel:
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
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line
More Bhagyashree Shahakar's questions See All
Similar questions and discussions