Cold Fusion Cloning fuses your PCR insert to a linearised vector using a proprietary enzyme blend, with no ligase or phosphatase step. A typical reaction takes around twenty minutes and gives you over 90 percent positive clones.
Cloning & Expression Systems
Cloning Tools and Kits
Cloning kits and specialised competent cells for expression-ready constructs
Getting your insert into an expression-ready vector means picking the right host strain, the right assembly chemistry and the right destination vector, in that order. Competent cells matched to your application come first. Cold Fusion Cloning handles ligation-free assembly in a single step. The PrecisionShuttle System moves your verified ORF between vectors.
Competent cells set your transformation efficiency. Chemically competent cells cover routine cloning. Reach for electrocompetent cells when you need highest efficiency, for example for building a library. The strain itself should match the project. Endura suits lentiviral cloning and CRISPR/Cas9 editing. Phage display strains handle antibody and peptide panning. ClearColi produces endotoxin-free protein. OverExpress strains take on toxic or membrane proteins. The strain notes list direct replacements for TOP10, DH5alpha, DH10B, JM109 and XL1-Blue.
Cold Fusion Cloning fuses your insert to a linearised vector, with no ligation and no phosphatase step. The PCR-amplified insert is mixed with the vector, incubated for five minutes at room temperature, then ten minutes on ice. That takes you to transformation-ready DNA in about twenty minutes. Typical reactions give over 90 percent positive clones. Any insert goes into any site in any vector, by hand or on an automated workflow. Your kit comes with competent cells included, and scales from ten reactions to a 96-well format for parallel projects.
The PrecisionShuttle System moves your sequence-verified ORF between vectors with a simple cut-and-paste digestion and ligation. It starts from a genome-wide TrueORF clone, covering human, mouse and rat genes, in the pCMV6-Entry or pCMV6-AC-GFP vector. Two rare-cutting enzymes, Sgf I and Mlu I, release the untagged ORF, so you can shuttle it into a mammalian, lentiviral or bacterial expression vector that shares the same cloning sites. Unlike Gateway cloning, it needs no recombination enzyme, carries no IP restrictions, and transfers ORFs up to 18 kb without the instability that affects larger Gateway inserts.
How to choose your cloning route
Your host strain comes first. Chemically competent cells cover routine transformations, and electrocompetent cells handle difficult inserts and library work.
Cold Fusion Cloning fits when you are building a new construct from a PCR insert and want a ligation-free, single-step reaction with a high success rate.
The PrecisionShuttle System fits when you already have a verified ORF and want to move it into a different expression vector without repeating the sequencing step.
Parent category: Cloning & Expression Systems
Applications
Endura competent cells clone unstable sequences reliably. Use them for lentiviral cloning and especially for cloning CRISPR gRNA Libraries as published in the Cold Spring Harbor protocol. Use them as your direct replacement for Stbl3 and SURE strains.
Phage display strains TG1, SS320 and E2738 give high transformation efficiency for antibody and peptide phage display libraries, with amber and non-amber options for your workflow.
ClearColi BL21(DE3) uses genetically modified lipopolysaccharide (LPS) to produce functionally clean, endotoxin-free recombinant proteins and plasmids. It suits your mammalian transfection, immunogenicity and toxicity assays, where clean protein matters most.
OverExpress C41(DE3) and C43(DE3) strains express toxic and membrane proteins that standard strains struggle with. The combo pack screens all four strains against your target.
Cold Fusion Cloning takes your PCR insert to transformation-ready DNA in about twenty minutes. Build expression or reporter constructs fast, without ligation or phosphatase steps.
The PrecisionShuttle System shuttles your sequence-verified TrueORF clone between mammalian, lentiviral and bacterial vectors by simple digestion, so a new tag needs no resequencing.
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Competent Cells
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Cold Fusion Cloning Kits
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PrecisionShuttle System
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Frequently asked questions
The PrecisionShuttle System uses two rare-cutting restriction enzymes, Sgf I and Mlu I, instead of a recombination enzyme. You need no licence to use it, and you can transfer ORFs up to 18 kb without the instability that recombination-based systems show above 4 kb.
Start with chemically competent cells for routine cloning. Choose electrocompetent cells when your insert is large, unstable, or destined for a library. Pick a strain built for your downstream use, such as Endura for lentiviral and CRISPR work or ClearColi for endotoxin-free protein.
Yes. Cold Fusion Cloning kits come with competent cells included, so you go from insert to transformed colonies with a single kit.
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