The DNA Shuffling Kit enables in vitro homologous recombination of DNA molecules, widely applied in molecular directed evolution. Starting from one or multiple homologous gene templates, the workflow includes DNase I random fragmentation, primer-free reassembly PCR and primer-mediated amplification PCR, to build libraries of chimeric and recombined gene variants.
Our kit is pre-formulated and ready-to-use. Researchers only supply purified homologous DNA templates, without preparing separate reaction components. The optimized protocol specifically favors homologous shuffling while suppressing unnecessary point mutations, saving heavy condition optimization work. It adapts to homologous target fragments around 1000 bp and supports up to 10 independent DNA shuffling reactions. This product is for research purposes only and cannot be used clinically.
Kit Components
| Component |
Specification |
| DNase I solution (1U/μL) |
10 μL |
| 10× DNA Shuffling Special Reaction Solution A |
100 μL |
| 10× DNA Shuffling Special Reaction Solution B |
100 μL |
| 2× Primer-free PCR MasterMix |
1 mL |
| 2× Primer-specific PCR MasterMix |
1 mL |
| Ultrapure water |
1 mL |
Storage & Shelf Life
Store at -20 °C; valid for 1 year.
Experimental Notes
- Template: Homologous DNA fragments (prepared by PCR or restriction digestion), total target length ~1000 bp. Total input DNA per reaction: 2–5 μg. Mixed homologous templates are recommended at equal mass ratio.
- Template purification: Gel purification is mandatory to eliminate residual plasmids, old primers and non-specific amplicons, which will disrupt shuffling.
- Preparations before reaction: Prepare mutated homologous gene fragments, second-round PCR primers and gel recovery kits in advance; preheat metal bath/water bath to 37 °C and 90 °C.
