📋 Workflow
Full workflow, products add-to-cart directly1
Seeding
30 minGrow to log phase, seed 24-well plate so 60-80% confluent at transfection
💡 Tip Optimize density per cell line
⚠️ Caution Use antibiotic-free medium during transfection
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2
Prepare transfection complex
15 minDilute lipid and nucleic acid in serum-free medium separately, mix and rest 15 min
💡 Tip Lipid:DNA ratio is key for efficiency
⚠️ Caution Do not dilute with serum, inhibits transfection
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3
Add to cells
10 minAdd complex dropwise, swirl to mix, incubate 4-6 h then replace medium
💡 Tip Medium change reduces toxicity
⚠️ Caution Handle gently to avoid detachment
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4
Expression detection
24–48 hContinue culture, observe reporter by fluorescence microscopy or verify by WB/qPCR
💡 Tip Include empty vector and untransfected controls
⚠️ Caution Transfection stress needs multiple controls
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Serum inhibits liposome transfection
- ② Unoptimized lipid:DNA ratio yields low efficiency or high toxicity
- ③ Improper confluency affects transfection rate
- ④ Excess reagent causes cell death
- ⑤ No reference plasmid to normalize efficiency
❓ FAQ
+How to verify success?
Check fluorescence for GFP tag and quantify by WB/qPCR.
+Low efficiency solutions?
Optimize reagent ratio, density, or switch to lentivirus/electroporation.
+siRNA vs plasmid?
siRNA for fast transient knockdown; plasmid for overexpression or stable lines.