Real lab failures, root causes, and fixes — curated and bilingually annotated by our team.
Primers that initially worked successfully for PCR gradually fail or produce poor results after storage
Very few or no colonies obtained when subcloning PCR products generated with primers containing restriction enzyme sites, despite successful amplification
PCR fails to amplify templates with high GC content or produces very low yields
Ethidium bromide-stainable material remains in the gel wells and does not migrate during electrophoresis
PCR produces a band of incorrect size or sequence not matching the expected target
No bands visible on gel electrophoresis after PCR amplification
Desired PCR fragment appears faint or barely visible on gel electrophoresis
Gel electrophoresis shows smeared or streaked bands instead of distinct sharp bands
Multiple unwanted bands appear on gel electrophoresis alongside or instead of the desired PCR product
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