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Product Overview
Limitless™ ELZ Fusion is a seamless, homology-based directional cloning kit that assembles fragments carrying 15–40 bp of vector homology in a single 37°C reaction, and that allows insertion of target genes at any position in any vector. The Limitless™ ELZ Fusion Seamless Cloning Kit is extremely simple to operate, requiring only linearization of the vector, mixing with PCR-amplified fragments that are completely homologous to the vector (15–40 bp) and reacting at 37°C, followed by direct transformation. The manufacturer states a positive clone rate of over 95%.
The kit supplies a 5× Limitless™ ELZ Fusion Mix for 25 reactions. A 20 µL reaction combines the linearized vector and the insert at an insert-to-vector molar ratio of 2:1 to 5:1 with 4 µL of 5× Mix and water, is incubated at 37°C for 30 minutes and then chilled on ice for 5 minutes. 10 µL of the reaction is used for transformation and the remaining 10 µL can be stored at 4°C or -20°C.
Specifications
| Format | Single 5× reaction mix |
|---|---|
| Cloning method | Seamless, homology-based directional cloning |
| Homology arm length | 15–40 bp |
| Insert-to-vector molar ratio | 2:1 to 5:1 |
| Reaction volume | 20 µL |
| Reaction time | 30 minutes at 37°C, then 5 minutes on ice |
| Positive clone rate | Over 95%, as stated by the manufacturer |
Kit Components
| Component | 25 rxn |
|---|---|
| 5× Limitless™ ELZ Fusion Mix | 100 µL |
Applications
- Seamless cloning of PCR-amplified fragments into linearized vectors
- Insertion of target genes at any position in a vector
- Insertion of multiple target genes into one vector
Storage & Handling
Stored at -20°C. The shelf life is 1 year. Shipping: Blue ice.
Usage Notes
- Design primers so that the amplified insert carries 15–40 bp of sequence completely homologous to the vector ends, and purify the insert before assembly.
- Linearize the vector by restriction digestion or PCR before setting up the reaction.
- Chill the reaction on ice for 5 minutes after the 37°C incubation.
- Store the unused 10 µL of the reaction at 4°C or -20°C.
Safety & Regulatory
For research use only. Not intended for clinical, diagnostic or therapeutic use. Follow the manufacturer's manual and standard laboratory safety practice.
Customization & Add-ons: Need this molecular biology reagent tailored to your workflow? We can help with bulk or custom pack sizes, alternative formats (spin column, magnetic bead or reagent-only kits; premixed or stand-alone reference dye), matched extraction, cDNA synthesis and qPCR workflows, and sourcing of related enzymes, ladders and consumables not in our catalog. Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services — our team will follow up with feasibility details and next steps.
Natural Product Alantolactone Targeting AKR1C1 Suppresses Cell Proliferation and Metastasis in Non-Small-Cell Lung Cancer. Frontiers in Pharmacology, 2022. 10.3389/fphar.2022.847906
MiR-143-5p regulates the proangiogenic potential of human dental pulp stem cells by targeting HIF-1α/RORA under hypoxia: A laboratory investigation in pulp regeneration. INTERNATIONAL ENDODONTIC JOURNAL, 2024. 10.1111/iej.14133
