No DNA Recovered from Plasmid Miniprep (DNA Cleanup & Plasmid Purification)

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No detectable plasmid DNA is recovered after miniprep purification. Column elution yields no nucleic acid by spectrophotometry or gel electrophoresis. Common causes: 1) Buffers added in incorrect order disrupting bind-wash-elute sequence 2) Ethanol not added to Plasmid Wash Buffer 2 before use 3) Plasmid loss during culture growth due to inadequate antibiotic selection 4) Bacterial culture lost selective pressure allowing plasmid-free cells to dominate Solutions: 1) Add buffers in correct sequence: resuspension → lysis → neutralization → wash → elution 2) Verify ethanol was added to Plasmid Wash Buffer 2 per kit instructions before first use 3) Use proper antibiotic at correct concentration to maintain selection (check plasmid resistance marker) 4) Confirm antibiotic stock solution is fresh and stored correctly (-20°C for most antibiotics)