Plasmid DNA Extraction Miniprep Protocol

critical

No DNA Recovered from Plasmid Miniprep

No detectable plasmid DNA is recovered after miniprep purification. Column elution yields no nucleic acid by spectrophotometry or gel electrophoresis.

💡 4 · ✓ 4

severe

Partial Digest with Multiple Uncut Bands

Gel shows mixture of fully digested, partially digested, and uncut DNA. Some recognition sites cleaved while others remain intact in the same molecule.

💡 5 · ✓ 5

severe

Low Plasmid Yield Due to Incomplete Lysis

Plasmid miniprep yields are significantly below expected levels. A260/280 ratio may be normal but total DNA concentration is low.

💡 5 · ✓ 5

severe

Mini-prep yields almost no plasmid DNA

Eluate DNA concentration is very low; diagnostic digest and gel show no detectable target band.

💡 4 · ✓ 4

severe

Incomplete Restriction Enzyme Digestion

Partial digestion observed with both expected fragments and uncut substrate visible on gel; incomplete linearization of plasmid DNA.

💡 6 · ✓ 6