A number of different methods, technologies, and protocols are available for performing the actual cloning reaction. To choose the one that best meets your needs, you will need to analyze the current parameters of your experiment and plan ahead for any downstream procedures. This section discusses several highly efficient cloning technologies:
- Restriction Enzyme Digestion & Ligation Cloning: is an industry standard for molecular biologists
- TOPO Cloning: provides simple, convenient reactions typically in less than 5 min
- Gateway Cloning Technology offers the greatest flexibility in vector choices and downstream applications
- Seamless Cloning and Assembly: Tools optimized to clone up to 4 DNA fragments simultaneously into virtually any linearized E. coli vector in a 30 minute room temperature reaction
- Type IIs Assembly: Clone up to 8 fragments simultaneously and without scars, and avoid rearrangements when cloning homologous or repetitive sequences
- GeneArt Gene Synthesis and Strings DNA Fragments: 100% accuracy and optimization of genes to maximize expression
The table below can help you navigate your way through these technologies.
| Restriction Enzyme Cloning | TA-Cloning | TOPO (TA, blunt, directional) | Gateway Cloning Technology | GeneArt Seamless Cloning | GeneArt Type IIs Assembly | GeneArt High-Order Genetic Assembly | |
|---|---|---|---|---|---|---|---|
| Fragments Cloned Simultaneously | 1 | 1 | 1 | Up to 4 | Up to 4 | Up to 8 | Up to 10 |
| Max Fragment(s) Size | Variable | 1-3 kb | <5 kb (<10 kb for XL-TOPO) | Variable | Up to 10 kb, max total size of 40 kb | Up to 10 kb, max total size of 13-20 kb | Up to 100 kb, max total size of 110 kb |
| Gene Shuttling Between Vectors w/o PCR or Restriction Enzymes | NO | NO | NO | YES | NO | NO | NO |
| Seamless (No Extra Sequences) | NO | NO | NO | NO | YES | YES | YES |
| Use Your Own Vector | YES | NO | NO | YES (may require conversion) | YES | YES | YES |
| Time to Clone Multiple Fragments | Days to Weeks | Not Possible | Not Possible | >4 Days | 1 hr | 1 hr | 3 Days |
| 4 Fragment Cloning Efficiency | NA | NA | NA | 30% - 85% | 75% | >90% | >90% |
| Web-Based Vector Design Tool | NO | NO | NO | NO | YES | YES | YES |
Browse for vectors using our Vector selection tool
For Research Use Only. Not for use in diagnostic procedures.