5 Cloning Shortcuts From Thermo Scientific

Streamline your workflow with these cloning tools

Here, at Thermo Fisher Scientific, we have tools to support your entire molecular cloning workflow. Cloning human, animal, plant, bacterial, or even non-coding genes? We've developed awesome, time-saving, and cost-cutting shortcuts to expedite your cloning experiments! Traditional cloning is time consuming and doesn't guarantee recombinant clones. Try our Top 5 Cloning Shortcuts and clone within hours and forget about false positives.

1. CloneJET PCR Cloning Kit

Do you want an all-inclusive cloning kit that comes with everything you need, including sequencing primers and a useful control PCR product? All you have to do is add your DNA fragment and provide competent cells for transformation. Our CloneJET kit is a positive selection system that yields 99% recombinant clones in 5 minutes! The positive cloning vector, pJET1.2/blunt eliminates the need for dozens of plasmid minipreps or expensive selection assays to identify recombinant clones, saving you hours of work.

2. FastDigest Restriction Enzymes

Prepping DNA inserts or digesting minipreps and using multiple restriction enzymes? What's better than 1 universal buffer to accommodate all your restriction enzymes and complete digestion in 5-15 minutes? Maybe the fact that we sell 176 FastDigest restriction enzymes that can do this! Double and multiple digestions will not be a hassle anymore because 1 buffer will work for all of them. Also, the FastDigest buffer allows for direct sample loading and is compatible with DNA modifying enzymes, making your life even easier! How about using the time you save with our FastDigest restriction enzymes and buffer to watch this video on just how fast these enzymes are.

3. FastAP Thermosensitive Alkaline Phosphatase

Your plasmid is linearized and now you have to dephosphorylate the ends to prevent recircularization during ligation – you definitely need an alkaline phosphatase (AP)! FastAP Alkaline Phosphatase is a novel AP, which is active in all Thermo Scientific restriction enzyme buffers and PCR buffers. It dephosphorylates all types of DNA ends (blunt, 5'- and 3'-overhangs) in 10 minutes at 37°C. And, no tedious enzyme removal is necessary prior to ligation because FastAP can be quickly inactivated at 75°C in 5 minutes.

4. Rapid DNA Ligation Kit

Efficient ligation is key to cloning success. The Rapid DNA Ligation Kit enables fast sticky-end or blunt-end DNA ligation in only 5 minutes at room temperature. The kit contains T4 DNA ligase and a specially-formulated rapid ligation buffer optimized for fast and efficient DNA ligation. The best part: the ligation product can be used directly* for transformation.

* If transforming by electroporation, ligation products must always be purified.

Ligation of sticky- and blunt-end DNA fragments with the Rapid DNA Ligation Kit. Blunt-end ligation: pUC19 DNA/SmaI dephosphorylated vector was ligated with Lamda DNA/PvuII fragment (2.3 kb). Sticky-end ligation: pUC19 DNA/PstI dephosphorylated vector was ligated with Lambda DNA/PstI fragment (2.1 kb). The efficiency of ligation was determined by transformation of E. coli XL1-Blue cells.

5. aLICator Cloning and Expression System

If you prefer ligation independent cloning you should explore our aLICator Cloning Kits. The aLICator System uses robust directional LIC cloning technology for PCR products to streamline and facilitate the process of insert cloning into the expression vector. This is a 15 minute procedure that yields greater than 95% recombinant clones without any restriction digestion or ligation steps! This kit can also be ordered with different His-tags to expedite protein purification at the end of your experiment.