Synthetic Biology/DNA Assembly Selection Chart
NEB® now offers several products that can be used for DNA Assembly. Use this chart to determine which product would work best to assemble your DNA.
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NEBuilder® HiFi DNA Assembly: Master Mix Cloning Kit Bundle for Large Fragments |
NEB Gibson Assembly®: Master Mix Cloning Kit |
NEBridge® Golden Gate Assembly: BsaI-HFv2 Kit BsmBI-v2 Kit |
USER™ Enzyme NEB #M5505 Thermolabile USER II Enzyme NEB #M5508 |
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| Properties | ||||
| Removes 5′ or 3′ End Mismatches | ★★★ | ★ | N/A | N/A |
| Assembles with High Fidelity at Junctions | ★★★ | ★★ | ★★★ | ★★★ |
| Tolerates Repetitive Sequences at Ends | ★ | ★ | ★★★ | ★★★ |
| Generates Fully Ligated Product | ★★★ | ★★★ | ★★★ | NR |
| Joins dsDNA with Single-stranded Oligo | ★★★ | ★★ | NR | NR |
| Assembles Low Amounts of DNA with High Efficiency | ★★★ | ★★ | ★★ | ★★ |
| Accommodates Flexible Overlap Lengths | ★★★ | ★★★ | ★ | ★★ |
| APPLICATIONS | ||||
| 2 Fragment Assembly (Simple cloning) | ★★★ | ★★★ | ★★★ | ★★★ |
| 3-6 Fragment Assembly (one pot) | ★★★ | ★★★ | ★★★ | ★★★ |
| 7-11 Fragment Assembly (one pot) | ★★★ | ★★ | ★★★ | ★★★ |
| 12-50+ Fragment Assembly (one pot) (1) | ★ | ★ | ★★★ | NR |
| Template Construction for In vitro Transcription | ★★★ | ★★★ | ★★★ | ★★★ |
| Synthetic Whole Genome Assembly | ★★★ | ★ | ★★★ | ★ |
| Multiple Site-directed Mutagenesis | ★★★ | ★★ | ★★ | ★★ |
| Library Generation | ★★★ | ★★★ | ★★★ | ★★ |
| Metabolic Pathway Engineering | ★★★ | ★★ | ★★★ | ★★★ |
| TALENs | ★★ | ★★ | ★★★ | ★★ |
| Short Hairpin RNA (shRNA) Cloning | ★★★ | ★★ | ★ | ★ |
| gRNA Library Generation | ★★★ | ★★ | ★ | ★ |
| Large Fragment (> 10 kb) Assembly | ★★★ | ★★★ | ★★★ | ★★ |
| Small Fragment (< 100 bp) Assembly | ★★★ | ★ | ★★★ | ★★★ |
| Use in Successive Rounds of Restriction Enzyme Assembly | ★★★ | ★ | NR | ★ |
(1) Please visit neb.com/GoldenGate for more information
| KEY | |
|---|---|
| ★★★ | Optimal, recommended product for selected application |
| ★★ | Works well for selected application |
| ★ | Will perform selected application, but is not recommended |
| N/A | Not applicable to this application |
| NR | Not recommended |
