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)
RNA Depletion for RNA-seq severe

No Depletion Due to Incorrect Probe Design Input

Percentage of reads mapping to the targeted sequence does not decrease after depletion treatment. Sequencing analysis shows target RNA remains at original levels.

💡 4 causes ✓ 5 fixes
RNA Depletion for RNA-seq severe

Probe Integrity Compromised During Storage or Synthesis

Target RNA depletion efficiency is significantly reduced or absent. Sequencing shows minimal reduction in targeted reads despite proper protocol execution.

💡 3 causes ✓ 4 fixes
RNA Depletion for RNA-seq severe

DNA Contamination Interferes with RNA Quantification and Depletion

RNA depletion shows poor efficiency with high residual target reads. RNA quantification may be inaccurate and inconsistent with expected values.

💡 3 causes ✓ 4 fixes
RNA Depletion for RNA-seq moderate

Chemical Contaminants Inhibit Probe Hybridization

Depletion efficiency is reduced or completely absent. Target RNA remains at high levels after treatment despite using validated probe designs.

💡 4 causes ✓ 5 fixes
RNA Depletion for RNA-seq severe

Improper Temperature Ramp During Probe Hybridization

Probe hybridization step was unsuccessful resulting in poor depletion. Target RNA removal is inconsistent or incomplete across replicate samples.

💡 4 causes ✓ 5 fixes
RNA Depletion for RNA-seq moderate

Non-Uniform Depletion Across Targeted Sequences

Depletion efficiency varies significantly across different targeted sequences. Some target regions show good depletion while others remain at high levels.

💡 4 causes ✓ 5 fixes
RNA Depletion for RNA-seq moderate

Genome Annotation Version Mismatch Between Design and Analysis

Target sequence used to design probes differs from the reference used to evaluate depletion. Depletion appears ineffective when analyzed against current genome annotations.

💡 4 causes ✓ 5 fixes