AffiNGS® DNA Damage Repair Kit
Optimized enzyme mix for repairing damaged DNA, especially from FFPE (formalin-fixed paraffin-embedded) samples.
Key Benefits:
- High repair efficiency for FFPE-derived DNA.
- Reaction buffer compatible with end repair/adenylation workflows.
- No further purification needed after using thermosensitive proteinase K.
- Direct integration into library construction workflows.
Components:
| Components | AFG-BCL-000001 (24 rxns) |
AFG-BCL-000002 (96 rxns) |
| Damaged DNA Repair Enzymes | 72 μL | 288 μL |
Storage:
- Storage: −20°C to −10°C.
- Shipping: Use dry ice or combination of dry ice and ice packs, maintain between −40°C to −20°C.
Protocol:
Important Notes:
-
Use only with severely damaged DNA such as FFPE samples.
- Not compatible with Cell-free DNA (cfDNA) – may cause random fragmentation.
- Not compatible with Tissue or high-quality genomic DNA – unnecessary and may not be effective.
1. Recommended DNA Input Amount:
| Sample Type | Suggested Input DNA (mass) | Notes |
| FFPE DNA | 100–500 ng |
Recommended range |
| 200–300 ng | Common working range | |
| Up to 1 µg | Higher input gives better libraries (if available) | |
| <100 ng | Possible for rare samples; expect lower yield |
2. Reaction Setup (on ice):
Compatible with ultrasonic-based fragmentation protocols.
| Reagent | Volume | Notes |
| Fragmented FFPE DNA | X μL | Adjust to desired input amount (see above) |
| End Repair Buffer (from your kit) | Y μL | As per your library kit's recommendation |
| End Repair Enzyme Mix | Z μL | As per your library kit's recommendation |
| Damaged DNA Repair Enzymes | 3 μL | Enhances repair of damaged FFPE DNA |
| Nuclease-free Water | Up to final volume (e.g., 60 μL) | Adjust to total reaction volume |
3. Mixing & Incubation:
- Mix thoroughly by pipetting.
- Briefly centrifuge.
- Incubate using a PCR thermocycler (lid temperature: 105°C):
| Temperature | Duration |
| 30°C | 30 min |
| 65°C | 30 min |
| 4°C | Hold |
4. Next Steps:
Proceed directly to:
- Adapter ligation
- Purification
- Library amplification
Follow your library prep kit's instructions for all subsequent steps.