Restriction Enzyme Cloning Support
Explore our “Getting Started” and “Troubleshooting” sections for solutions to top inquiries and common problems. Browse through our "Guides and Tools" section to access a comprehensive portfolio of product-related support resources.
- I cannot transform my cells right away. Can I store my ligation reaction? If so, at what temperature should I store it?
- What is the difference between ExpressLink™ T4 DNA Ligase and other T4 DNA ligases?
- I have an older set of enzymes from your company that uses REact buffers. Are these buffers and enzymes interchangeable with the new enzymes?
- For how long do I digest my vector?
- What is the best ratio of insert:vector to use? Is there an equation to calculate this?
- I would like to perform a double digest with two different enzymes but the buffers are not compatible.
- I am doing restriction cloning and the enzyme I am using seems to be cutting non-specifically.
- I have cloned my gene into my vector and then transformed into TOP10 cells. I then did a plasmid miniprep followed by digestion of the DNA with Xba I. However, the vector is not cutting correctly.
- I am trying to clone an insert that is supposedly pretty toxic. I used DH5α and TOP10 cells for the transformation and got no colonies on the plate. Do you have any suggestions for me?
Guides and Tools
- Overview: Restriction Enzyme Digestion and Ligation
- Protocol: Cloning A-tailed PCR Fragments
- Selection Table: Which Cloning Method is Right for You?
- Selection Table: Recommended Universal Buffers for Double Digestion
- Selection Table: Relative Activities of Restriction Enzymes in Each Reaction Buffer and Buffer Compositions
- Supplemental Protocol: Rapid Ligation Protocol
Gene synthesis outperforms traditional cloning by providing fast turnaround times and guaranteed sequence accuracy. Construct design and optimization services are also available in order to fine tune the expression of your gene(s) of interest.
If you have a favorite cloning vector or a vector that you wish to use for a specific assay, let our TOPO® Cloning experts adapt it for you. Any vector you choose can be TOPO® adapted for high efficiency cloning of PCR products in 5 minutes.
GeneArt® Plasmid Services include flexible offerings covering construction of plasmid vectors customized for your needs, and subcloning of your sequence into any vector, including the Gateway® vector system. Concentrate on your research goals and leave your plasmid construction and subcloning work to us.
For Research Use Only. Not for use in diagnostic procedures.