In-Fusion Cloning—Data from the Field
Free of the limitations that come with traditional cloning methods, In-Fusion Cloning is unique in its ability to provide streamlined, seamless cloning for every experiment. Each system is a complete, fully optimized set of reagents that provides a high out-of-the-box success rate, even for difficult cloning projects. Take advantage of fast, directional cloning without constraints—clone any insert(s) into any vector at any locus.
- Overcome the limitations imposed by restriction site availability
- Clone with large inserts or vectors (up to 46-kb cosmids)
- Add multiple inserts to a single vector with one reaction
- Incorporate insertions, deletions, or point mutations
Exceptional accuracy is maintained across all cloning applications, regardless of vector type or insert composition. Unlike ligation-based methods, cloning success remains high even as vector size, insert size, or number of fragments increases. Subcloning is unnecessary and background levels are extremely low, reducing both screening time and the number of reactions required to obtain your final construct.
Take a look at the field tester data below to see how others have used this technology to spend less time cloning and more time advancing their research.
Cloning Without Ligase: A Fast, Accurate Alternative
Ti-Yu Lin, Graduate Student, University of Wisconsin
Madhusudan Rajendran, Research Assistant, University of Wisconsin—Madison
- Where multiple previous ligation-based cloning experiments had failed, In-Fusion Cloning generated positive clones on the first attempt, even with GC-rich or large inserts.
- Ti-Yu Lin, Graduate Student, University of Wisconsin
Beyond Ligase: More Efficient Single- and Multiple-Insert Reactions
Dr. Samuel Bru, Postdoc, Orphan Cyclins Group, Universitat Internacional de Catalunya
- In-Fusion Cloning and ligation were tested side-by-side in reactions with one, two, and three inserts. In-Fusion technology far outpaced ligation, showing impressive cloning efficiency, even with multiple-insert reactions.
- Dr. Samuel Bru, Postdoc, Orphan Cyclins Group, Universitat Internacional de Catalunya
Fast, Direct Cloning into Large Expression Vectors
Ansul Lokdarshi, Graduate Research Assistant, University of Knoxville, TN
- A large binary vector for in planta localization studies of YFP-tagged proteins was constructed with just one cloning reaction, bypassing subcloning and achieving a success rate of 100%.
- Ansul Lokdarshi, Graduate Research Assistant, University of Knoxville, TN
Overcoming Restriction Site Limitations with Lentiviral Vectors
Jun Yang, Instructor, University of Texas Medical Branch
- In-Fusion Cloning was used to solve the problem of limited restriction site availability in lentiviral vectors. Five different inserts were directly cloned into the final vector with a success rate nearing 100%.
- Jun Yang, Instructor, University of Texas Medical Branch
Outperforming the TOPO Cloning Workflow with Speed and Accuracy
Juanita Von Dwingelo, Ph.D. Candidate, University of Louisville
- Researchers cloning proteins of interest for yeast two-hybrid studies found that In-Fusion Cloning provided better, faster results than those obtained with TOPO cloning methods, including a cloning accuracy of 100%.
- Juanita Von Dwingelo, Ph.D. Candidate, University of Louisville
Solving a Synthesis Challenge with Easy Multiple-Insert Cloning
Christian Joerg Braun, Postdoc, MIT
- When secondary structure prevented the synthesis of an activation domain, In-Fusion technology was used to clone the full-length sequence by inserting two overlapping fragments into an 11-kb Cas9 expression vector.
- Christian Joerg Braun, Postdoc, MIT