Incorrect Reaction Efficiency Due to Oligo Binding to Non-Molecular Biology Tubes (PCR (Sigma Guide))

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Variable and incorrect standard curve efficiency when using serial dilutions; effect more pronounced when same dilution series stored at 4°C and reused; inconsistent differences between amplification plots; problem resolves when different operator uses different tubes Common causes: 1) Template oligonucleotides binding to non-molecular biology grade plastic tubes 2) Loss of template through adsorption to tube walls during storage 3) Use of inappropriate plasticware without low-retention properties 4) Extended storage at 4°C exacerbates binding effects Solutions: 1) Switch to molecular biology-grade, low retention plasticware (e.g., Eppendorf 1.5 mL tubes) 2) Prepare fresh dilution series immediately before use 3) Avoid storing dilutions for extended periods, especially at 4°C 4) Verify all plasticware used is suitable for molecular biology applications 5) Use certified low-binding tubes for oligonucleotide and template storage