The Gibson Assembly® HiFi 1-Step Kit

Eliminate time consuming restriction cloning, the need to subclone after TA cloning, and avoid recombination-based cloning that adds ‘att sites’. Get the clone you want in the vector you want. Insert multiple fragments in a single round of cloning and never subclone again! The Gibson Assembly® HiFi 1-Step Kits provide a simple and effective alternative to seamlessly construct synthetic genes, genetic pathways, as well as entire genomes. This simple one tube, one step, isothermal approach allows researchers to create large recombinant DNA constructs, with multiple inserts, in 1 hour.

Gibson Assembly® HiFi 1-Step technology utilizes a special mix of enzymes optimized to assemble the DNA quickly and efficiently. Unlike other approaches, SGI-DNA’s Gibson Assembly process utilizes Phusion® High-Fidelity DNA polymerase for 3’ end extension. This results in higher numbers of seamless error free clones ready for multiple downstream applications.

The Gibson Assembly® HiFi 1-Step Kit is available as either a 2X master mix or 4X high concentration (HC) master mix. The 4X Gibson Assembly HiFi HC 1-Step kit requires only 2.5 µL of mix per 10 µL reaction making it ideal for assemblies using less concentrated DNA samples. Using either the original HiFi 1-Step 2X format or the HiFi HC 1-Step 4X format, multiple DNA fragments can be joined with greater than 90% cloning efficiency. Even large constructs up to 100kb in length can be efficiently assembled in multi stage reactions.

Summary

  • Fast and efficient — Complex assemblies of > 90% cloning efficiencies in 1 hour with less than 5 minutes hands-on time
  • Restriction Digest Free — Clone without the need for restriction enzyme sites
  • Optimal for fragment sizes ranging from 500bp to 32kb
  • Utilize any vector using simple design strategies — avoid subcloning
  • Multiple Inserts – Assembles up to 5 inserts at one time
  • Large Constructs — create constructs up to 100kb in length in multi stage assemblies
  • Cloning Specificity — Enhanced cloning specificity of PCR products due to homology matching between amplicon and vector
  • Adaptability — Easily adapted for multiple DNA manipulations, ss oligonucleotides and assembling repetitive sequences
  • Proven published method — Since 2009, the Gibson Assembly® method has been cited by more than 1,200 scientific publications
Gibson Assembly HiFi 1-Step Kit

How It Works

Gibson Assembly® method employs an innovative blend of enzymes in a single tube reaction in a 1 hour isothermal reaction. The process begins by designing dsDNA fragments with 20 – 40bp overlapping ends. The DNA fragments are mixed with the Gibson Assembly® mix for assembly.

  • 5' chew back
  • DNA fragments anneal
  • Phusion® DNA polymerase extends 3’ ends
  • Any nicks are sealed

The resulting construct is a double stranded, fully sealed, and error free molecule that can be used immediately for numerous molecular biology applications such as direct transformation, PCR, RCA, and other molecular biology applications.

Applications

  • Multiple Insert Cloning – Clone multiple inserts into a vector without relying on the availability of restriction sites
  • Large Fragment Assemblies – Assemble gene clusters and genome-size fragments
  • Mutagenesis – Site-directed mutagenesis to make simultaneous changes in a single reaction
  • See now the team at JCVI created the first synthetic cell using Gibson Assembly®
  • Application Notes

Gibson Assembly® HiFi 1-Step Provides High Cloning Efficiency

Gibson Assembly HiFi 1-Step Kit Cloning Efficiency Chart
Gibson Assembly® HiFi 1-Step delivers high colony output with up to 98% cloning efficiency for 2 fragment assembly (4.2kb) and up to 95% for 5 fragment assembly (5.9kb).

Gibson Assembly® HiFi 1-Step Reagents (2X)

Products Catalog # Price
Gibson Assembly® HiFi 1-Step Starter Kit GA1100-S $80
Gibson Assembly® HiFi 1-Step Kit, 10 rxn GA1100-10 $159
Gibson Assembly® HiFi 1-Step Kit , 50 rxn GA1100-50 $630
Gibson Assembly® HiFi 1-Step Master Mix (2X), 10 rxn GA1100-10MM $149
Gibson Assembly® HiFi 1-Step Master Mix (2X), 50 rxn GA1100-50MM $580
Gibson Assembly® Positive Control GA1000-05C $63

Gibson Assembly® HiFi HC 1-Step Reagents (4X)

Products Catalog # Price
Gibson Assembly® HiFi HC 1-Step Starter Kit GA1100-4XS $82
Gibson Assembly® HiFi HC 1-Step Kit, 10 rxn GA1100-4X10 $163
Gibson Assembly® HiFi HC 1-Step Kit , 50 rxn GA1100-4X50 $648
Gibson Assembly® HiFi HC 1-Step Master Mix (4X), 10 rxn GA1100-4X10M $153
Gibson Assembly® HiFi HC 1-Step Master Mix (4X), 50 rxn GA1100-4X50M $597
Gibson Assembly® Positive Control GA1000-05C $63

Bulk Packaging

Gibson Assembly® HiFi 1-Step MasterMix (2X)

MasterMix Volume Product Catalog # Total # of reactions Standard Price Discount Discounted Price Price/Reaction
5 mL Gibson Assembly® HiFi 1-Step MasterMix (2X) GA1100-B05 1000 $11,600 56.90% $5,000 $5.00
10 mL GA1100-B10 2000 $23,200 61.21% $9,000 $4.50
20 mL GA1100-B20 4000 $46,400 65.52% $16,000 $4.00
30 mL GA1100-B30 6000 $69,600 69.83% $21,000 $3.50
40 mL GA1100-B40 8000 $92,800 74.14% $24,000 $3.00
50 mL GA1100-B50 10000 $116,000 78.45% $25,000 $2.50

Gibson Assembly® HiFi HC 1-Step MasterMix (4X)

MasterMix Volume Product Catalog # Total # of reactions Standard Price Discount Discounted Price Price/Reaction
3 mL Gibson Assembly® HiFi HC 1-Step MasterMix (4X) GA1100-4XB03 1200 $14,328 57.87% $6,180 $5.15
5 mL GA1100-4XB05 2000 $23,880 61.18% $9,270 $4.64
10 mL GA1100-4xB10 4000 $47,760 65.49% $16,480 $4.12
15 mL GA1100-4XB15 6000 $71,640 69.81% $21,630 $3.61
20 mL GA1100-4XB20 8000 $95,520 74.12% $24,720 $3.09
25 mL GA1100-4XB25 10000 $119,400 78.43% $25,750 $2.58

Above mentioned pricing is valid only in the U.S. For international pricing please contact customerservice@sgidna.com.

Prices do not include shipping and handling. Submit by 2pm (PST) to receive order by the next day. For bulk packaged size, lead time is ~3 weeks.

Gibson Assembly method is also available under commercial license, please inquire at info@sgidna.com

Place Your Order

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Email: customerservice@sgidna.com

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