📋 Workflow
Full workflow, products add-to-cart directly1
RNA extraction and QC
2 hExtract high-quality total RNA, check concentration, purity and integrity (RIN)
💡 Tip RIN ≥7 usually acceptable for sequencing
⚠️ Caution RNase degradation affects results
🧪 Materials for this step (click to shop by spec/brand)
2
Library construction
1 dayDeplete rRNA or enrich mRNA, reverse-transcribe to cDNA, ligate adapters and amplify
💡 Tip Choose ribodepletion per species
⚠️ Caution Control cycles to avoid PCR bias
🧪 Materials for this step (click to shop by spec/brand)
3
Sequencing
1–3 daysQuantify, pool libraries and sequence on high-throughput sequencer
💡 Tip Read depth per sample count
⚠️ Caution Pool evenly for uniformity
🧪 Materials for this step (click to shop by spec/brand)
4
Data analysis
2 daysQC, align, quantify and perform differential expression and enrichment analysis
💡 Tip Multiple-testing correction prevents false positives
⚠️ Caution Correct batch effects
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① RNA degradation lowers RIN
- ② High rRNA residual wastes sequencing
- ③ Excess PCR introduces expression bias
- ④ Batch effects mask true differences
- ⑤ Low depth misses lowly-expressed genes
❓ FAQ
+How to set RNA-seq depth?
Depends on goal; deeper for low-expression or novel transcripts (20-50M/sample).
+Ribodepletion vs poly-A enrichment?
poly-A enriches mRNA standard; ribodepletion retains ncRNA more comprehensively.
+How to correct batch effects?
Balance batches in design and correct with methods like ComBat.