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Help Understanding PCR Troubleshooting ? Unexpected Band in

 
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eremystery



Joined: 07 Feb 2025
Posts: 1
PostPosted: Fri Feb 07, 2025 7:42 am    Post subject: Help Understanding PCR Troubleshooting ? Unexpected Band in Reply with quote
Hi everyone,

I'm running a standard PCR reaction, but I'm getting an unexpected extra band in my gel electrophoresis results. My target amplicon is supposed to be around 500 bp, but I'm consistently seeing an additional faint band at approximately 700 bp.

I've tried:
Reducing the annealing temperature slightly
Running a no-template control (which showed no contamination)
Using fresh primers and reagents
Has anyone encountered something similar? Could it be primer-dimer, nonspecific amplification, or possibly genomic DNA contamination? Any troubleshooting tips would be really appreciated!

Thanks in advance!
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