Choosing a Cloning Method
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:
The table below can help you navigate your way through these technologies. |
| Restriction Enzyme Cloning | TA-Cloning | TOPO® (TA, blunt, directional) | Gateway® | GeneArt® Seamless Cloning | GeneArt® High-Order Genetic Assembly | |
|---|---|---|---|---|---|---|
| Fragments Cloned Simultaneously | 1 | 1 | 1 | Up to 4 | Up to 4 | Up to 10 |
| Max Fragment (s) Size | Variable | 1-3 Kbp | <5 kbp (<10 Kbp for XL-TOPO®) | Variable | Up to 10 Kbp, max total size of 13 Kbp | Up to 100 Kbp, max total size of 110 Kbp |
| Gene Shuttling Between Vectors w/o PCR or Restriction Enzymes | NO | NO | NO | YES | NO | NO |
| Seamless (No Extra Sequences) | NO | NO | NO | NO | YES | YES |
| Use Your Own Vector w/o Modification | YES | NO | NO | YES | YES | YES |
| Use Pre-existing Fragments w/o Modifications | NO | NO | NO | NO | YES (1 Fragment) | YES (3 Fragments) |
| Time to Clone Multiple Fragments | Days to Weeks | Not Possible | Not Possible | >4 Days | 1 Day | 3 Days |
| 4 Fragment Cloning Efficiency | NA | NA | NA | 30% - 85% | 75% | >90% |
| Web-Based Vector Design Tool | NO | NO | NO | NO | YES | YES |
| List of Available Vectors | GO | GO | GO | GO | GO | GO |
