Automated Plasmid Purification

Automated plasmid purification is an instrument-driven, walkaway workflow that performs lysis, binding, washing, and elution to enable high-yield, high-purity plasmid DNA. Using magnetic bead-based technology, researchers can boost sample throughput, increase reproducibility of results, and reduce contamination and hands-on time. Plasmid DNA purification is a critical step in applications including molecular biology, cell and gene therapy, and mRNA vaccine development. Traditional plasmid prep methods are often manual, slow, and inconsistent, and require centrifugation, pipetting, and multiple wash steps that increase hands-on time and reduce reproducibility.

As labs scale up higher throughput and plasmid isolation prep sizes, the move from manual plasmid purification to automated plasmid purification systems has become important. Automation enables faster, cleaner, and more quality results—reducing variability and freeing scientists to focus on discovery instead of repetitive tasks. Thermo Fisher Scientific offers a complete plasmid purification automation ecosystem featuring two advanced technologies:

• MagMAX Pro HT NoSpin Plasmid MiniPrep Kit—a fast, magnetic bead-based plasmid DNA isolation kit for small-scale and high-throughput applications
• KingFisher PlasmidPro Maxi Processor—a fully automated, maxi-scale plasmid DNA purification instrument that automates numerous steps from lysis to elution with a single prefilled cartridge

These solutions help provide transfection-grade and endotoxin-free plasmid DNA respectively for downstream applications such as cloning, gene editing, AAV production, and mRNA synthesis with speed, reliability, and scalability at every stage.

Request demo   Plasmid eBook

There are several advantages of automating plasmid DNA purification, including helping save time, improving efficiency, and increasing reliability of results. The MagMAX Pro HT NoSpin Plasmid Miniprep Kit along with KingFisher instruments and the KingFisher PlasmidPro Maxi Processor Endotoxin-Free Cartridge along with the KingFisher PlasmidPro Maxi Processor offers a range of key benefits for mini and maxi-scale plasmid DNA purification. Enhance your productivity and performance in plasmid DNA purification with these user-friendly bench-top solutions.

Back to top

"We are pioneering the development of programmable mRNA therapeutics, which hold the promise of more personalized and targeted treatments. Accelerating research and discovery is crucial for advancing the field of genetic medicine and ensuring that therapies are both effective and safe for specific tissue and cell types. 

The Applied Biosystems MagMAX Pro HT NoSpin Plasmid MiniPrep Kit has been instrumental in our research due to its high throughput capabilities and consistent, reliable yields that have helped significantly streamline our plasmid DNA purification process. Not only has this kit enhanced our workflow efficiency, but also has empowered our team to focus on the innovative aspects of our research, accelerating our journey towards delivering groundbreaking mRNA therapies." 

- Verified customer

The KingFisher PlasmidPro Maxi Processor is designed to simplify the entire plasmid purification process with automation, creating a 1-step process of loading your culture and pressing “Start run.” Starting from the overnight bacterial culture to obtaining purified plasmid DNA in under 75 minutes, the instrument allows researchers to load their culture and walk away with less than 5 minutes of hands-on time. Improve productivity without compromising quality. Free your scientists from laborious, time-consuming manual plasmid purification methods. This benchtop system yields advanced transfection-grade plasmid DNA of <0.1 EU/µg, suitable for a range of applications including transfection, protein expression, and gene therapy, among other sensitive applications from discovery to development. Scientists in biotech, biopharma, research institutions, service labs, and beyond—grow your culture, and this instrument will handle the plasmid purification with push-button simplicity.

Back to top

This workflow shows how to use the MagMAX Pro HT NoSpin Miniprep plasmid purification on the KingFisher Flex system or KingFisher Apex system. Steps include adding reagents to the 96 deep-well plates, preparing the sample plate, selecting the correct extraction protocol on the KingFisher instrument, starting the run, and following the prompted steps to complete the run. Obtain the elution plate with purified pDNA about 35 minutes after the start of the run cycle.

The KingFisher PlasmidPro system offers a streamlined and user-friendly workflow for plasmid purification. Its simple design and intuitive interface make it easy to operate, even for users with minimal training. With automated steps and precise control, the KingFisher PlasmidPro system can help simplify the entire process, from sample loading to elution, helping ensure reliable and efficient plasmid purification every time.

Back to top

The KingFisher PlasmidPro system consistently achieves high pDNA concentrations, exceptional elution volumes, and expected total yields, helping ensure reliable and reproducible results. Additionally, the system produces desired A260/A280 and A260/A230 purity ratio values, and preserves a high percentage of supercoiled form, indicating the integrity of the purified DNA. With its exceptional performance, the automated system is an excellent solution for researchers seeking efficient and high-quality maxi-scale plasmid DNA purification.

Figure 1. Gel image and supercoiled determination of purified pDNA. (A) Representative samples from Plasmid C (6.6 kb) in lanes 1–15 were purified using the KingFisher PlasmidPro instrument and compared to pDNA from a manual column-based kit in lanes 16, 17, and 18 run in 1% agarose gel. L1 is an Invitrogen linearized dsDNA ladder and L2 is the supercoiled pDNA ladder. The gel image shows that the supercoiled pDNA is the predominant form at 6.6 kb. (B) Densitometry analysis was performed with purified pDNA samples run on an agarose gel using the iBright Imaging and Analysis Software. Data represents the percent supercoiled pDNA from a subset of samples analyzed using the KingFisher PlasmidPro Maxi Processor. The box plot represents the distribution of data. Each dot represents an individual purification process. N is the number of purified samples from each plasmid size. The results show an average supercoiled pDNA at >80% for all three plasmid sizes. The percent supercoiled is comparable to the manual process for each plasmid B, C, and D at 88%.

Figure 2. Efficiency of transfection with plasmids purified on the KingFisher PlasmidPro Maxi Processor relative to manually purified plasmids. Plasmids B and C were used to transfect HuH7 mammalian cells. The y-axis values indicate transfection efficiency relative to the pDNA isolated using the manual purification process (set at 100% for both plasmids). The data represent reporter gene expression after transfection with 100 ng of pDNA isolated using SEAP expression from plasmid B and luciferase expression from plasmid C were compared to those of pDNA purified from the same overnight bacterial cultures using the column-based manual purification process. The box plots show the distributions of the data, with each dot representing an individual purification.

The magnetic bead technology of the MagMAX Pro HT NoSpin Plasmid Miniprep Kit helps deliver improved performance over traditional anion exchange and silica membrane technologies (Figure 3a). Plasmid DNA purified using the MagMAX Pro HT NoSpin Plasmid Miniprep Kit has a significantly higher transfection efficiency than column-based plasmid purification methods (Figure 3b). This allows for a wide range of downstream applications, such as transfection into sensitive cell lines, preparation of short hairpin vectors, enzymatic modifications, cloning, restriction digestion, sequencing, and in vitro transcription/translation.

Figure 3. Consistent yields and efficient transfection with MagMAX Pro HT NoSpin Plasmid Miniprep Kit. In Figure 3a, with respect to yields and isolation chemistry, these isolations were carried out using the KingFisher Apex instrument. Two different vectors of varying sizes have been isolated, pSEAP2 (5.2 kB) and PNM (11.4 kB). pSEAP2 was transformed into Top10 cells, cultured in LB,  and gave an average yield of 18.75 µg pDNA for an number of 384 samples.  pNM was transformed into DH5 alpha, cultured in Terrific Broth, giving a yield of 22.61 µg. Figure 3b shows transfection efficiency of plasmid DNA isolated using the MagMAX Pro HT No Spin Plasmid Miniprep Kit compared to traditional column-based Purification of plasmid DNA. S1 , S2, and S3 , represent three independent randomized samples from a single 96 well plate run. pDNA was isolated from Top10/pSEAP2 culture using MagMAX Pro HT and a Silica Filter column control.

Back to top

Back to top