QuikChange Lightning by Agilent Technologies on Transcriptic

Generate single point mutations quickly and easily with QuikChange Lightning from Agilent on Transcriptic


QuikChange kits provide a highly efficient non-PCR method for reliable site-directed mutagenesis. The QuikChange method uses a linear amplification strategy with only the parental strand serving as the DNA template. Combining this with the highest fidelity polymerases leads to a significant reduction in unwanted second-site errors, thus avoiding complications and downstream delays in screening and analysis.

The kit has been optimized for mutagenesis of plasmids of up to 14 kb, allowing rapid, efficient, and accurate mutagenesis of small and large plasmids with a single kit. Exclusive to the QuikChange Lightning Site-Directed Mutagenesis Kit is a proprietary Pfu-based polymerase blend and the newly optimized Dpn I enzyme, which together allow for mutagenesis in approximately one hour, plus an overnight transformation.

The reagent performance you expect, with the scalability of Transcriptic

Figure 1 - A screenshot of the experiment configuration screen. Uploaded a CSV of mutagenic primers and set your experimental parameters. It's as easy as that to generate 100's of mutants.

Using QuikChange Lightning on Transcriptic is as easy as configuring  5 parameters and clicking run. This enables users to generate anywhere from 3 mutants up to 96-well plates with ease. This ease of scalability can enable you to probe a much larger parametric space of protein sequence-space to investigate effects of clinical mutations or engineer better proteins.

The protocol takes care of ordering the mutagenic oligonucleotides, performing the mutagenesis, bacterial transformation, bacterial culture, plate spreading and finally colony picking. Following this, you can use your picked colonies in any experiments you like such as growth curves or sequencing all on Transcriptic or have the mutants shipped back to your lab.


Validated on Transcriptic

The QuikChange site-directed mutagenesis kit has been implemented on the automated Transcriptic platform and validated with the kit controls and test samples. Both figures 2 and 3 indicate the Sanger sequencing results of 2 adjacent mutated sites of a CA to GG. 3 of the 4 picked and sequenced colonies we found to be positive for the desired mutations.

Figure 2 - Sanger sequencing of 4 picked colonies from the experiment showed the 2 adjacent point mutations were achieved in 3 out for the 4 colonies selected.

Figure 3 - Inspection of the sequence chromatograms showed that the reads taken for the successful mutants were of high quality.

GET STARTED GENERATING 100’S OF MUTANTS TODAY

To get started with QuikChange Lightning and Transcriptic, log into your Transcriptic account and click “Launch Run”, from the protocol browser select the QuikChange Lightning protocol and configure it for your experiment. Make sure to have the template DNA to mutate in your inventory as well as a list of mutagenic primers to make your mutants.

 

Learn more

Agilent QuikChange Lightning Homepage

Agilent QuikChange Lightning user manual


What is Transcriptic?

Transcriptic is the robotic cloud laboratory for life sciences. Transcriptic’s revolutionary, automated cellular and molecular biology platform enables scalable life science research with flexibility and precision. The convenient and simple web interface allows biologists to control their science and generate data from virtually anywhere in the world. Transcriptic frees researchers to focus on the creative aspects of their science by providing faster, more reliable and repeatable processes with vastly lower capital costs than previously possible. More information on Transcriptic can be found at www.transcriptic.com.