Why i can´t see a good band in electrophoresis gel?

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Zhou Wei

your plasmids should be digested by restriction endonuclease first and then examined with agarose gel electrophoresis.

Ander Hernández Plágaro

I totally agree with Zhou Wei, before running your plasmid in an agarose gel you should digest it with a restriction enzyme of your choice (e.g., EcoRI, HindIII, BamHI). If you don't do so, plasmid coiling will make it migrate unevenly and you will get extra signals. Take into account that if your plasmid harbours more than one restriction site for your chosen enzyme, you will see an extra band for each extra restriction site.

It's also recommended to run undigested samples as controls to check if the plasmids present any anomaly.

Good luck!!

M. R. Mozafari

Hi,

Please have a look at the following PDF files (available online) and web links:

PDF Troubleshooting Guide for DNA Electrophoresis

res.hmu.edu.iq/Portals/0/Users/Bazhdar/DNA_Troubleshooting.pdf

Troubleshooting Guide for DNA Electrophoresis. 3.7. Gel shift effect. DNA electrophoresis problem. 1. Low intensity of all or some. DNA bands. 2. Smeared DNA. ...

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electrophoresis Methods, Protocols and Troubleshootings

www.protocol-online.org/biology-forums/electrophoresis.html

Help troubleshoot SDS gel - smear down well - (reply: 2); High concentration DNA on spectrophotometer but no band on agarose gel. WHere is - DNA ... MoBio Plasmid Purification and Gel Extraction Kits - Problems with MoBio Kits (reply: 3) ...

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Agarose Gel Electrophoresis Tips & Tricks | Thermo Fisher Scientific

https://www.thermofisher.com/us/en/.../agarose-content-with-tips-and-tricks.html

A higher agarose percentage enhances resolution of smaller bands; conversely, ... The type of buffer used to run DNA in agarose gel electrophoresis depends ...