NEB recommends NEB 10-beta Competent E. coli , High Efficiency (NEB #C3019) for assemblies larger than 15 kb. NEBuilder® HiFi DNA Assembly Bundle for Large Fragments offers a discount for purchasing these competent cells with the NEBuilder HiFi DNA Assembly Master Mix.
NEBuilder HiFi DNA Assembly Master Mix was developed to improve the efficiency and accuracy of DNA assembly. This method allows for seamless assembly of multiple DNA fragments, regardless of fragment length or end compatibility. This method has been used to assemble either single-stranded oligonucleotides or different sizes of DNA fragments with varied overlaps (15–80 bp). It has utility for the synthetic biology community, as well as those interested in one-step cloning of multiple fragments due to its ease of use, flexibility and simple master-mix format.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|NEB® 10-beta/Stable Outgrowth Medium||4||1 X|
|NEB® 10-beta Competent E. coli (High Efficiency)||-80|
|NEBuilder® HiFi DNA Assembly Master Mix||-20|
|NEBuilder® Positive Control||-20||2 X|
|pUC19 Vector||-20||0.05 ng/µl|
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