Gibson Assembly® HiFi 1 Step Kit, Synthetic Genomics

Supplier: Synthetic Genomics
GA1100-50 GA1100-10 GA1100-S
10820-608EA 124.17 USD
10820-608 10820-610 10820-614
Gibson Assembly® HiFi 1 Step Kit, Synthetic Genomics
Nucleic Acid Reagents Cloning Kits and Ligation
Generate seamless plasmid DNA or BAC clones without restriction digestion

The Gibson Assembly process begins by designing dsDNA fragments with 20 – 40bp overlapping ends. For the Gibson Assembly HiFi 1-Step reaction, these DNA fragments are combined with a 2X Gibson Assembly® HiFi 1-Step Master Mix and incubated. This during this time, complementary ends are generated, annealed, and sealed. This this one-step, isothermal process can be performed in less than 60 minutes and results in a transfection or transformation ready, double stranded, fully ligated DNA construct.

The GA 1-Step Master Mix (2X) contains a proprietary mixture of enzymes and reagents optimized to facilitate one-step assembly of double standed DNA fragments. This mastermix includes a proof-reading polymerase that mediates junction repair resulting assembled constructs with low rates of junction errors and high sequence fidelity. The GA Positive Control (2X) is sufficient for 2 reactions (5 and 10 reaction kits) or 5 reactions (50 reaction kit). This control consists of a mixture of 10 ng of a 1.5 kb insert and 30 ng of a 2.7 kb vector containing an ampicillin resistance gene. Selection for the 4.2 kb assembled construct can be performed using LB agar plates with 100 μg/ml ampicillin, 0.1 mM IPTG, and 40 μg/ml X-Gal.

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® HiFi 1-Step Kit, the simultaneous cloning of one to five inserts with sizes ranging from 500 bp to 32kb can be accomplished in a single tube, in a single step, using a robust isothermal reaction that yields cloning efficiencies of greater than 90%.

Información para pedidos: Kits with 5, 10, or 50 reactions are available. All kits contains one vial of GA 1-Step Master Mix (2X) and one vial of GA Positive Control (2X) and a detailed instruction manual.
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