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Real mistakes other researchers hit, and how to get them right.
Massive cell death after transfection — reagent toxicity is too high
Why it fails Transfection reagent dose is too high or inherently toxic
No amplification — Ct value missing or > 40
Why it fails Template (RNA / cDNA) degraded or concentration too low
Mycoplasma Contamination - Silent Growth Inhibitor
Why it fails Introduction through poor aseptic technique (mycoplasma found on human skin)
Confirmed colony later shows recombination or deletion
Why it fails Insert contains repeat sequences, LTRs, or other unstable elements that recombine
No bands at all (blank membrane)
Why it fails Transfer failed or membrane transfer was incomplete
No colonies at all — total failure
Why it fails Seeding density was too low (or no live cells survived)
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Restriction Digest Analysis
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Enhanced Genetic Analysis of Single Human Bioparticles Recovered by Simplified Micromanipulation from Forensic ‘Touch DNA’ Evidence
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Mouse Hindbrain Ex Vivo Culture to Study Facial Branchiomotor Neuron Migration
Robot-Assisted Radical Antegrade Modular Pancreatosplenectomy Including Resection and Reconstruction of the Spleno-Mesenteric Junction
Experimental Manipulation of Body Size to Estimate Morphological Scaling Relationships in Drosophila
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