📋 Workflow
Full workflow, products add-to-cart directly1
Column and first nucleoside
30 minChoose a synthesis column pre-loaded with 3' first nucleoside for solid-phase synthesis
💡 Tip Column scale matches synthesis amount
⚠️ Caution Keep anhydrous conditions
🧪 Materials for this step (click to shop by spec/brand)
2
Stepwise coupling cycles
HoursRepeat deprotection-coupling-oxidation cycles per sequence to extend stepwise
💡 Tip Coupling efficiency relates to purity
⚠️ Caution Reagents must be fresh and anhydrous
🧪 Materials for this step (click to shop by spec/brand)
3
Deprotection and cleavage
2–4 hDeprotect and cleave from support with base like ammonia, remove base protecting groups
💡 Tip RNA needs special deprotection
⚠️ Caution Optimize temp/time per sequence
🧪 Materials for this step (click to shop by spec/brand)
4
Purification and QC
HoursPurify oligo by PAGE or HPLC, verify length and purity by mass spec
💡 Tip Quantify by OD260
⚠️ Caution Long/modified oligos need rigorous purification
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Incomplete coupling gives deletion sequences
- ② Moisture/degraded reagent lowers yield
- ③ Missed phosphorothioate/base
- ④ Insufficient purification contaminates
- ⑤ RNA degrades unless RNase-free
❓ FAQ
+How to quantify synthesized primers?
Dissolve, read OD260, convert to concentration/moles by extinction coefficient.
+Long oligo low purity?
Use PAGE or IP-RP-HPLC rigorous purification to remove deletions.
+Can modified oligos be synthesized?
Yes; phosphorothioate, fluorescent, biotin and 2'-modifications via corresponding monomers.