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.


NEBuilder®
HiFi DNA
Assembly
NEB #E2621
NEB #E5520
NEB #E2623
Gibson Assembly®
NEB #E5510
NEB #E2611
NEB Golden
Gate Assembly Mix
NEB #E1600
USER™ Enzyme
NEB #M5505
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        
Simple Cloning (1-2 Fragments) ★★★ ★★★ ★★★ ★★★
4-6 Fragment Assembly ★★★ ★★★ ★★★ ★★★
>6 Fragment Assembly ★★★ ★★ ★★★ ★★★
Template Construction for In vitro Transcription ★★★ ★★★ ★★★
Synthetic Whole Genome Assembly ★★★
Multiple Site-directed Mutagenesis ★★★ ★★ ★★ ★★
Library Generation ★★ ★★ ★★ ★★
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


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