Failure Case Library

Real lab failures, root causes, and fixes — curated and bilingually annotated by our team.

All Techniques (7) Nucleic Acid Quantification (9) Lipid Transfection (8) End-point PCR Primers (9) Cell Culture (Contamination) (7) PCR (Sigma Guide) (12) Flow Cytometry (Controls) (8) PCR / qPCR Plastics (9) PCR (Invitrogen Guide) (8) Cell Culture (Precipitates) (6) Cell Culture (Cell Death) (8) ELISA (Sigma Guide) (6) PCR / RT-PCR Amplification Problems (5) Flow Cytometry (Paraformaldehyde Fixation) (14) Flow Cytometry (Sample Considerations) (14) ELISA (R&D Guide) (10) Western Blot Immunodetection (19) Flow Cytometry (CST Guide) (8) Immunohistochemistry (CST Guide) (14) Immunoprecipitation (CST Guide) (14) ChIP (CST Guide) (8) ELISA Development (9) Western Blot (Sigma Protocol) (8) IP-Western Blot (6) Western Blot (CST Guide) (8) Flow Cytometry (Fixation Buffers) (7) Flow Cytometry (Isotype Controls) (7) Flow Cytometry (Fixation & Permeabilization) (9) Flow Cytometry (Fc Blocking) (7) Flow Cytometry (Compensation) (7) Flow Cytometry (Autofluorescence) (7) Flow Cytometry (Troubleshooting) (8) Tissue Imaging (Autofluorescence) (9) ELISPOT (8) Immunoprecipitation (Protein Obstruction) (1) Immunoprecipitation (No Protein Detected) (5) Immunoprecipitation (High Background) (8) Immunoprecipitation (High Antibody Elution) (1) Immunohistochemistry (No Staining) (9) Immunohistochemistry (High Background) (9) ChIP (PCR Amplification Problems) (4) ChIP (Low Signal) (8) ChIP (Low Resolution with High Background) (6) ChIP (High Background) (1) Western Blot (Blue Background) (1) Western Blot (Unusual Gel Band Appearance) (3) Western Blot (Unexpected Multiple Bands) (7) Western Blot (Misshapen / Uneven Bands) (5) Western Blot (Bands at Wrong MW) (5) Western Blot (Detection Problems) (7) Western Blot (Weak / No Signal) (6) ELISA (Standard Curve Fit Problems) (6) ELISA (Inconsistent Results / High CV) (6) ELISA (High Background) (8) ELISA (Signal Problems) (11) ELISA (Competitive) (18) Cell-free DNA Extraction (8) Bacterial rRNA Depletion (4) RNA Depletion for RNA-seq (7) LAMP (Loop-mediated Isothermal Amplification) (7) HMW DNA Extraction (Monarch) (7) NGS Library Preparation (NEBNext Ultra II) (7) RNA Cleanup (4) DNA Cleanup & Plasmid Purification (7) PCR (Polymerase Chain Reaction) (26) Restriction Enzyme Digest (13) Western Blot (9) Immunohistochemistry (IHC) (6) Colony Formation Assay (10) Transwell Migration / Invasion Assay (5) qPCR (RT-qPCR) (5) Transfection (8) Plasmid Construction (1) Sanger Sequencing (2) Plasmid Mini-prep (1) Plasmid Construction (Double Digest) (1) Protein Extraction (4) Transfection (Co-transfection) (1) Transfection (siRNA Knockdown) (1) CCK-8 Cell Viability Assay (5) Wound Healing Assay (5)
LAMP (Loop-mediated Isothermal Amplification) severe

LAMP Reaction Orange Before Amplification Starts

The colorimetric LAMP reaction appears orange instead of pink/magenta prior to incubation at 65°C, indicating premature pH drop that compromises amplification.

💡 4 causes ✓ 5 fixes
LAMP (Loop-mediated Isothermal Amplification) critical

No-Template Control Turns Yellow (Contamination)

The no-template control (NTC) reaction turns yellow after incubation at 65°C, indicating contamination with amplifiable target nucleic acid or positive control template.

💡 4 causes ✓ 6 fixes
LAMP (Loop-mediated Isothermal Amplification) severe

Internal Control Fails Due to Sample pH Incompatibility

The internal control reaction fails to turn yellow after incubation at 65°C when spiked into sample, though it works in water, indicating sample matrix interference.

💡 4 causes ✓ 5 fixes
LAMP (Loop-mediated Isothermal Amplification) moderate

Internal Control Fails with Non-Human Sample Source

The internal control reaction fails to turn yellow when added to nucleic acid isolated from non-human sources (e.g., animal tissue, environmental samples), as the control targets human-specific sequences.

💡 3 causes ✓ 5 fixes
LAMP (Loop-mediated Isothermal Amplification) severe

Internal Control Fails Due to RNA Template Degradation

The internal control reaction fails to amplify when using RNA samples, while DNA-based controls work normally, indicating RNA-specific quality issues.

💡 5 causes ✓ 6 fixes
LAMP (Loop-mediated Isothermal Amplification) critical

Positive Control Remains Pink After Incubation

The positive control reaction fails to turn yellow following 30-minute incubation at 65°C, indicating no amplification occurred despite presence of target template.

💡 5 causes ✓ 5 fixes
LAMP (Loop-mediated Isothermal Amplification) moderate

Amplification Failure Due to Inadequate Reaction Mixing

Reactions show inconsistent color change or partial amplification with some replicates failing, caused by reagent stratification or incomplete mixing before incubation.

💡 4 causes ✓ 5 fixes