📋 Workflow
Full workflow, products add-to-cart directly1
Fragmentation
30 minFragment purified nucleic acids to desired size by enzymatic or sonication methods
💡 Tip Fragment size determines read length and output
⚠️ Caution Fragment uniformly without degradation
🧪 Materials for this step (click to shop by spec/brand)
2
End repair and A-tailing
1 hRepair ends and add A-tail to blunt ends for adapter ligation
💡 Tip End repair quality affects ligation efficiency
⚠️ Caution Avoid over-processing causing sample loss
🧪 Materials for this step (click to shop by spec/brand)
3
接头连接与条码
1 hLigate sequencing adapters (with barcodes), purify to remove unligated adapters
💡 Tip Barcodes index multiple samples
⚠️ Caution Avoid adapter/amplicon cross-contamination
🧪 Materials for this step (click to shop by spec/brand)
4
Amplification and QC
2 hPCR-amplify the library, purify and QC by quantification (qPCR/site) and Bioanalyzer
💡 Tip Control cycles to avoid PCR bias
⚠️ Caution Accurate quantification is key for pooling
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Degraded nucleic acid yields poor library
- ② Adapter contamination gives low-complexity data
- ③ Excess PCR cycles introduce bias
- ④ Inaccurate quantification harms pooling uniformity
- ⑤ Cross-contamination causes barcode crossover
❓ FAQ
+How to judge library quality?
Check concentration, single clean size peak without adapter peak, adequate qPCR on-target.
+Required sequencing depth?
Varies: RNA-seq 20-50M/sample, WGS 30× genome.
+Can libraries be stored long-term?
Store short-term at -20°C, sequence promptly, avoid repeated freeze-thaw.