Gibson Assembly® ULTRA Kits and Master Mixes (2X)

Supplier: Telesis Bio
Gibson Assembly®
GA1200-50MM GA1200-B05 GA1200-10 GA1200-50 GA1200-10MM GA1200-S
10820-792EA 130.9 USD
10820-792 10820-790 10820-794 10820-652 77463-576 10820-618
Gibson Assembly® ULTRA Kits and Master Mixes (2X)
Nucleic Acid Reagents Cloning Kits and Ligation
Generate seamless plasmid DNA or BAC clones without restriction digestion.

  • Maximum number of fragments: 10
  • Cloning efficiency: 95% Full-length clones
  • Insert size range: 100 to 100000 bps
  • Total construct size range: 1000000 bps
  • Reaction time: 80 minutes

The Gibson Assembly process begins by designing dsDNA fragments with 20 – 40bp overlapping ends. For the Gibson Assembly Ultra reaction, a two-step process is used. First, DNA fragments are combined with a 2X Gibson Assembly® Ultra Master Mix A. Incubated to generate overlapping free ends. After enzyme inactivation the reaction is slowly cooled to anneal overlapping ends. Next the 2X Gibson Assembly® Ultra Master Mix B is added to allow the generation of a fully sealed construct. This this highly efficient and robust cloning method takes about 80 minutes and results in a transfection or transformation ready, double stranded, fully ligated DNA construct.

GA Ultra Assembly reaction occurs in two steps using the addition of two master mixes in two sequential steps. In the first step, the GA Ultra Master Mix A (2X) creates single-strand DNA 5’ overhangs by chewing back DNA from the 3’ end. This reaction is cooled under conditions that allow for annealing of complementary overlap regions. Next, the GA Ultra Master Mix B (2X) is added. During this step, nucleotides are incorporated into the construct to fill in the gaps in the annealed DNA fragments and nicks are sealed to create a contiguous DNA construct.

The Gibson Assembly method allows the insertion of one or more linear double stranded DNA fragments into a virtually any vector without the need to rely on compatible restriction sites. Gibson Assembly is significantly faster than traditional restriction enzyme digest-based cloning and proven for the cloning of both small and large double stranded DNA fragments. Using the Gibson Assembly® Ultra Kit, the simultaneous cloning of one to fifteen inserts with sizes ranging from 100 bp to 100 kb can be accomplished in a single tube using a highly efficient two-step reaction that yields cloning efficiencies of 95%.
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