Hi, a gel electrophoresis analysis (1% agarose) was performed after restriction enzyme analysis to test if the insert is inside the plasmid. The restriction enzymes were supposed to cut once inside the insert and once inside the vector. On the first gel, smear is observed in the gel. Could someone help me find out what's the reason of this? (on wells 8,10 accidentally ladder was pipetted so those can be ignored). this was done for around 55 minutes at 100V.

The gel was repeated because of this and similar results were observed in the second gel. The smear was there again. More loading dye was used in the second gel (2ul) so maybe this could be a reason? This gel was ran at 120V for 45minutes.

I would really appreciate the help. Thank you.