I'm looking at primers at different annealing temperatures and my gel is showing two distinct streaking bands at the top of the ladder and at the bottom of the ladder. I'm using DNA from ferns within the pgiC gene.
Very first reason for appearance of smear can be
-the gel is not poured correctly, it will not solidify evenly, thus causing the molecules to smear.
2) Due to excessive loading or overloading of wells and also if the gel is moved after the sample is placed in the well, it can cause the sample to spill out of the well that causes smear to appear.
3)May be due to incorrect buffer system or temperature changes or inappropriate voltage supplied
4)Avoid contamination: DNA contaminated with protein causes smear to appear.
I think it is a common problem with Gel Electrophoresis when you go through PCR.
5)Lastly: Your DNA might be degraded during gel extraction, so when you run the extracted DNA you only get smear.
Good Luck!!!
Very first reason for appearance of smear can be
-the gel is not poured correctly, it will not solidify evenly, thus causing the molecules to smear.
2) Due to excessive loading or overloading of wells and also if the gel is moved after the sample is placed in the well, it can cause the sample to spill out of the well that causes smear to appear.
3)May be due to incorrect buffer system or temperature changes or inappropriate voltage supplied
4)Avoid contamination: DNA contaminated with protein causes smear to appear.
I think it is a common problem with Gel Electrophoresis when you go through PCR.
5)Lastly: Your DNA might be degraded during gel extraction, so when you run the extracted DNA you only get smear.
Good Luck!!!
the top of the gel may be genomic dna and the bottom may be one primer extending on its own because the other primer cannot anneal.Impossible to say without a picture.If this is a pcr please give cycling conditions and primer sequences and post a picture
I agree with Paul. The upper band may be your DNA template. For the bottom band it may be dimers of primers. you could see that if your PCR did'nt work.
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