Complete DNA Extraction to Gel Electrophoresis Protocol

severe

Low Gel Extraction Yield Due to Improper Gel Dissolution

DNA yield from gel extraction is poor. Undissolved agarose particles may be visible or column flow is impeded.

💡 4 · ✓ 4

severe

Low or No Amplification Due to Template Issues

PCR yields little to no product visible on gel electrophoresis. Expected band is absent or extremely faint despite using standard reaction conditions.

💡 6 · ✓ 6

severe

PCR Inhibition from Contaminated Template

No or weak PCR amplification despite correct reaction setup, with template DNA containing residual contaminants from extraction or purification that inhibit polymerase activity.

💡 4 · ✓ 6

moderate

No Band or Faint Band Due to Template Quality Problems

PCR fails or produces weak products despite correct reagent concentrations and cycling parameters, due to compromised template DNA quality or presence of inhibitors.

💡 4 · ✓ 4

moderate

Smeared Bands Due to Template Quality or Concentration Issues

Gel displays smeared bands suggesting degraded products or heterogeneous amplification. Template-related issues such as degradation, contamination, or excessive concentration are suspected.

💡 6 · ✓ 6

moderate

Nonspecific Amplification and Smearing on Gel

Gel shows multiple bands, smears, or high background instead of single clean product band. May include primer-dimers at bottom of gel.

💡 6 · ✓ 6