GenBuilder® DNA Assembly
GenScript's GenBuilder® DNA Assembly technology takes recombination-based cloning to the next level. Pushing the bounds of innovation, the GenBuilder® technology allows for seamless fusion of one to twelve gene fragments with the highest efficiency. Whether using fragments longer than 5 kb, unpurified PCR products, single stranded DNA inserts, or have only fifteen minutes to complete the cloning reaction- the GenBuilder® DNA Assembly technology helps you build your constructs quickly and accurately.
GenBuilder® perfectly pairs with GenScript's GenParts® DNA Fragments service. This service enables synthesis of your gene fragments (100-2,000 bp) with delivery in as few as 2 days. The DNA fragments thus generated can be cloned directly into any vector of your choice using the GenBuilder® DNA Assembly.
Seamless cloning made easy as 1-2-3!
Short assembly time
DNA Assembly works with unpurified PCR fragments
Incubation in the enzyme mix for only 15 minutes
Highest cloning efficiency in the industry
Rigorously tested among 8 DNA assembly kits
Quality control by assembling 12 fragments with >90% positive rate
Performance verified by >1,000 reactions per day in production line
- I routinely receive 400ng of GenParts® fragment. Is this sufficient for the GenBuilder® cloning?
For a 1 kb fragment, 400 ng GenParts® fragment is generally sufficient for about six GenBuilder® reactions.
- Should the GenParts® fragment be gel-purified or spun-purified prior to using it in the GenBuilder® reaction?
All GenParts® fragments are gel-purified before shipping, so no additional purification is required before using them with the GenBuilder® cloning kti.
- What is the minimum required overlap length for DNA assembly with GenBuilder?
A 40 bp overlap is optimum for obtaining the maximum cloning efficiency. For cloning one or two fragments into a construct, a 15 bp overlap may be sufficient. For larger number of fragments, we recommend using a 40 bp overlap to increase cloning efficiency. Please see experiment 4.1 (pages 5-6) to see data for cloning together 2 fragments with different over>lap lengths.
- When using ssDNA as a bridge, how long can the ssDNA be (not including the overlap site)?
We have tested 20 bp ssDNA inserts without issues. For longer ssDNA fragments, the acceptable insert length may have to be determined experimentally.
|How GenBuilder® DNA Assembly works|
1. Prepare the DNA fragments with overlaps and the linearized vector for assembly.
2. Add DNA fragments and linearized vectors in the GenBuilder® master mix. Mix well and incubate at 50°C for 15 minutes.
3. Transform assembly product into competent cells and plate on selective medium. Obtain colonies containing the assembled DNA.
GenBuilder® Kit features
#1 You choose your samples, GenBuilder® DNA Assembly delivers the results
- Directional cloning of DNA fragments may be achieved using recombinant arms 15-40 bp in length.
- DNA inserts may be generated by PCR or gene synthesis.
- The vector may be linearized by any restriction enzyme of the user's preference.
- Up to six DNA fragments may be cloned together into a vector in a single-step reaction using GenBuilder® Cloning kit. Up to 12 fragments ,may be cloned together using GenBuilder® Plus.
- DNA fragments as long as 8 kb may be cloned.
- The PCR products may be used for cloning directly without purification.
- DNA Assembly is completed in as few as 15 minutes.
#2 Best on the market, GenBuilder® DNA Assembly outcompetes all other DNA Assembly kits with performance
- For multiple cloning scenarios such as cloning five gene fragments into a single vector, using unpurified PCR products, and enzyme performance after repeated freeze-thaw cycles, the GenBuilder® DNA Assembly outperforms all other kits for DNA assembly.
#3 GenBuilder® DNA Assembly- a single solution for multiple cloning applications
- Vector generation by one-step single and multi-gene cloning.
- Gene mutation (addition, deletion and point mutation of base pairs) using ssDNA. Codon saturation mutagenesis using ssDNA as a bridge.
- High throughput cloning, and much more.
- Restriction site independent
- One-step cloning of 12 fragments
- Seamless assembly
- Suitable for long fragments (> 8 kb)
|Features||CloneEZ® PCR Cloning kit||GenBuilder® Cloning Kit||GenBuilder® Plus Cloning Kit|
|1||Unique need fulfilled by the kit||Cloning one fragment at room temperature||Multi-gene cloning for up to 6 fragments||Multi gene cloning for up to 12 fragments|
|2||Cost per reaction||$7.30||$7.90||$9.80|
|3||Number of fragments that can be cloned into a linearized vector||1||6||12|
|4||Cloning efficiency (insert was cloned into the vector) %||80%||>90%||>90%|
|5||Plasmid library construction||Not suitable||Suitable||Recommended|
|6||Minimum time taken for cloning the maximum number of inserts in a vector||30 min||15 min||15 min|
|7||Cloning unpurified PCR product||No||Efficient||Highly efficient|
|8||Assembly with ss oligos (single stranded oligos)||No||No||Yes|
|9||Can be used for high throughput cloning||Yes||Yes||Yes|
|11||Positive control||Linearized pUC57 with one positive control fragment||Linearized pUC57 with two fragments to reconstitute RFP for visual determination of cloning efficiency||Linearized pUC57 with two fragments to reconstitute RFP for visual determination of cloning efficiency|