📋 Workflow
Full workflow, products add-to-cart directly1
RNA extraction and QC
2 hExtract total RNA including small RNAs, check concentration, integrity and purity
💡 Tip Small RNAs need dedicated extraction to retain
⚠️ Caution RNase contamination degrades small RNAs
🧪 Materials for this step (click to shop by spec/brand)
2
Reverse transcription
2 hReverse-transcribe miRNA with stem-loop primers, or lncRNA by poly(A)/oligo-dT
💡 Tip Choose tailing/stem-loop strategy per target
⚠️ Caution Short small RNAs need specialized primers
🧪 Materials for this step (click to shop by spec/brand)
3
qPCR quantification
2–3 hQuantify by qPCR with specific primers/probes, normalize to reference RNAs
💡 Tip Use suitable reference (e.g. U6)
⚠️ Caution Choose reference per RNA type
🧪 Materials for this step (click to shop by spec/brand)
4
Analysis and target validation
2 hCalculate relative expression by 2^-ΔΔCt, validate ncRNA regulatory targets
💡 Tip Multiple references improve reliability
⚠️ Caution Target regulation needs functional validation
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Small RNAs lost in routine extraction
- ② RNase contamination causes degradation
- ③ Wrong reference biases results
- ④ Non-specific primers amplify wrongly
- ⑤ Degradation/plateau distorts quantification
❓ FAQ
+miRNA vs gene quantification difference?
miRNAs are short; need stem-loop or tailing RT and length-specific probes.
+How to choose reference?
U6/RNU6B for miRNA, GAPDH/ACTB for lncRNA, verify stability.
+How to validate ncRNA targets?
Combine prediction databases with reporter, mutagenesis and overexpression/knockdown experiments.