Pro Ligation-Free Cloning Kit (20 reactions)
Cat. No. | E086 |
Name | Pro Ligation-Free Cloning Kit (20 reactions) |
Unit | 20 Reactions |
Category | Molecular Biology Enzymes and Kits |
Description |
The Pro Ligation-Free Cloning Kit allows for the robust assembly of multiple DNA fragments in one step. Using a ligation-independent cloning method, as many as six DNA fragments are assembled seamlessly without concern for restriction enzyme site availability. abm’s Pro Ligation-Free cloning is a simple yet highly versatile method to rapidly clone highly complex, multiple component constructs. Key Kit Features:
Testimonial: "I tried the molecular cloning of inserting two DNA fragments (2Kb & 0.5Kb) into a 4Kb backbone by using the kit. After the transformation, I got about 100 colonies, and when I picked 24 colonies for colony PCR, the bright band from gel running told me that I got 20 potential hits, which is awesome. I sent some plasmids for sequencing, the result I received just now unveiled they are carrying the exact right fragment insertion. Collectively, this is a powerful kit." - Jianfeng Lan, The Buck Institute |
Application |
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Storage Condition |
Store at -20°C |
Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. E086 |
Do I need to purify the PCR product first before use with this kit? Or can I use low melting agarose for the PCR gel and just melt the band portion and use in the kit? | |
The kit can be used in either scenario.
1. The kit comes with Cloning Optimizer and Assembly Enhancer enzymes which allow the user to not have to do gel extraction/purification following the amplification of DNA fragments, as methylated template DNA and single stranded DNA (e.g. primers) will be removed. 2. If PCR reactions contain multiple non-specific bands, it is highly advisable to either optimize the PCR or perform gel extraction, as the contaminating bands will be present in the cloning reaction if not prior removed. |
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