4 replies to this topic

[mimilka]

I have run PCR in 8 different temperatures and I run it on a gel. However instead of bands I got circles? Any suggestions what it can be? Is it possible the concentration of my genomic DNA or primer concentration was too high? And why they are at different positions if it was the same sample?

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[perneseblue]

mimilka, on Mar 21 2009, 12:46 PM, said:

I have run PCR in 8 different temperatures and I run it on a gel. However instead of bands I got circles? Any suggestions what it can be? Is it possible the concentration of my genomic DNA or primer concentration was too high? And why they are at different positions if it was the same sample?

I will have to say, this gel was run rather poorly. What is the expected band size? Are there suppose to be 8 lanes with PCR products in them? I would suggest that run the gel again.

[pcrman]

To me, they do not look like bands at all. They are probably just dirty spot in the gel because they are so round in shape.

[WOW]

pcrman, on Mar 22 2009, 11:08 AM, said:

To me, they do not look like bands at all. They are probably just dirty spot in the gel because they are so round in shape.

I agree with dirty spot. And it seems the gel was not well prepared, since even the marker was not clear. Try more before make the conclusion. May it help you and good luck.

[bochum]

Next time cook the agarose properly and if your adding Etbr directly into the gel mix it properly....this big spot may be undissolved Etbr or unmelted agarose. And i dont think it is ur pcr product because first spot is almost under the marker lane rather than below first lane....