Many automated liquid handling systems and purfication instruments move reagents into and out of wells to perform the different steps of nucleic acid isolation. In contrast, Thermo Scientifc KingFisher Sample Purification Systems use permanent magnetic rods with disposable tip combs to collect beads from solution, then release them into successive wells, each containing reagents for the next step of isolation. This highly effective method of bead collection and transfer leads to clean extractions, efficient results, and preservation of sample. Using a simple process (bind, wash, elute), KingFisher instruments can automate the extraction of any analyte of interest that can be bound to a bead, even from challenging samples.
Moving beads instead of liquids results in higher purity because no non-target organic matter or contaminants from liquid reagents are transferred from well to well. It also minimizes sample loss and results in higher yield since you can optimize your protocol so that no beads, with their target nucleic acids attached, are left behind in solution.
Manual sample preparation and sample loss can introduce inconsistencies into samples, which show up in your data as outliers and wider error bars. High-end downstream applications such as qPCR, NGS, digital PCR, and mass spectrometry can be very sensitive to these inconsistencies. KingFisher instruments reduce user processing errors while increasing the reproducibility of your results. Our data show that samples processed with KingFisher instruments have consistent results across different runs and different users.
KingFisher instruments help save you valuable time by removing manual steps and reducing overall processing time when compared with manual preparation or even with other sample preparation instruments. This not only improves workflow efficiency but also reduces exposure to potentially infectious samples.
With KingFisher automation, each step in your workflow—such as reagent, antibody, target capture, washes, and elution—is represented by a single plate in the instrument carousel (or, for the KingFisher Duo Prime, rows of wells in the plate). Once the plates are prepared and loaded, simply press Start and the instrument carries out the programmed protocol automatically. As each step is completed, the carousel rotates to bring the next plate underneath the magnetic head and tip combs.
A KingFisher Flex system can isolate viral RNA from a 96-well plate in only 25 minutes. A typical lab using 1–4 Flex instruments can easily process 1,000–4,000 samples in an 8-hour day. Using highly automated KingFisher Presto instruments can process the same number of samples but with far less attention from laboratory staff.
KingFisher systems can gently and efficiently isolate and purify nucleic acids, proteins, and cells from a range of sample types for a wide variety of downstream applications.
Different chemistries handle a multitude of sample types. KingFisher systems offer the flexibility to process even difficult samples such as blood, saliva, mucus, seawater, soil, fecal matter, fish scales, and other sample types. KingFisher systems can handle sample preparation for an extraordinary range of applications.
Different sample types and applications require different chemistries to extract the analyte. To simplify selection and ease of use, we’ve developed kits and solutions that contain optimized beads, buffers, and protocols to maximize purity and yield for each application and sample type. The Kits, Reagents, and Protocols section contains details about these applications and kits, but here are a few highlights:
MagMAX kits for optimized nucleic acid extraction. For automated extraction of DNA and RNA using KingFisher systems, the reagents of choice are Applied Biosystems MagMAX kits. For example, MagMAX Viral/Pathogen Nucleic Acid Isolation Kits can be used to extract and purify RNA from swabs or other samples types. Other popular MagMAX kits help you extract genomic DNA (gDNA) from a variety of samples; circulating cell-free DNA for liquid biopsies; DNA and RNA from formalin-fixed, paraffin embedded (FFPE) samples for clinical cancer research; and total RNA and miRNA for gene expression research.
Dynabeads and SMART Digest reagents for other targets. KingFisher instruments can use Invitrogen Dynabeads magnetic beads or Thermo Scientific SMART Digest kits to capture and isolate other nucleic acids, proteins, and cell structures. For example, you could use a SMART Digest kit to automate protein digestion of a monoclonal antibody like ipilimumab or rituximab (Rituxan®) for in-depth peptide mapping and characterization. You can pair a KingFisher instrument with Dynabeads magnetic beads to capture CD9+ tumor-derived exosomes for downstream analysis with flow cytometry or western blot.
All kits and many reagents include modifiable protocols optimized for KingFisher instruments. To find kits and reagents for your application, browse the Kits, Reagents, and Protocols tab or use our Selection Guide. If you need assistance customizing reagents or developing protocols for your particular application needs, our Field Application Specialists and Technical Sales Specialists will be happy to help.
KingFisher sample preparation instruments are optimized for use with MagMAX nucleic acid purification kits, Dynabeads reagents, and KingFisher plates and plastics. These products are designed and manufactured to exacting standards and the protocols developed for them have been tested, refined, and validated. When you use these products and protocols together, you can have confidence in the performance of your instrument and the purity, yield, and sample integrity of your results.
KingFisher systems are optimized to work with MagMAX kits, Dynabeads reagents, and KingFisher plates and plastics.
Optimization for Dynabeads magnetic bead reagents (which are also included in MagMAX purification kits) is central to consistent binding and reliable results. Dynabeads are superparamagnetic particles, meaning that they exhibit magnetic properties when placed in a magnetic field, with no residual magnetism once removed from the field. They are gentle on cells and molecules, capturing targets directly from biological samples in tubes or plates, without columns or centrifugation.
View our not a legal edit video to learn why micro-sized Dynabeads magnetic beads are better for studying natural cell processes than nano-sized magnetic “microbeads.” (7:21)
Dynabeads’ tightly controlled manufacturing processes produce magnetic beads of uniform spherical shape, size, and smoothness. With the same surface area offered for each experiment, you can achieve more reliable and reproducible results.
Dynabeads magnetic beads have a defined surface to carry out the necessary binding, with no inner surface to trap unwanted proteins. A. Dynabeads products are the most uniform, monodispersed superparamagnetic beads, manufactured with highly controlled product quality to help ensure the highest degree of reproducibility. B,C,D. Magnetic particles from alternative suppliers have variable shapes and sizes that trap impurities, resulting in lower reproducibility and increased nonspecific binding.
Within this optimization, KingFisher systems are flexible, giving you the freedom to modify and customize the workflow to the requirements of your experiment or process. The instruments themselves offer:
To help ensure that KingFisher systems can adapt to your changing research needs, Thermo Scientific BindIt BindIx software is included free with all KingFisher instruments.
The software lets you create, modify, and store your own protocols on your PC, transfer them to your KingFisher instrument, and track their execution on the instrument screen during the course of your run. It also allows you to define and edit steps and parameters for every part of your protocol—bind, wash, and elute—as well as define KingFisher plates and reagents in the plate layout. If you have already developed a manual protocol, you can convert to an automated protocol in minutes!
Protocols are available for MagMAX and Dynabeads nucleic acid isolation kits, as well as Dynabeads and SMART Digest protein purification products. Once loaded onto your instrument, you can modify the protocols and store them for future reuse.
Streamlined protocols for SARS-CoV-2 testing from nasopharyngeal swab samples were just as efficient at isolating four viral RNA targets as the standard protocol.
KingFisher systems are designed to be easy to install, learn, and use. In most cases, you’ll be ready to run in 10 minutes or less. The instruments are also customizable to your lab’s requirements.
Workflows are simple, too. Once set up, simply scroll to the protocol you want to run on the instrument screen and follow the step-by-step instructions on how to prepare and load each plate. Then just press Start and watch it happen! (Or walk away as the instrument follows the protocol automatically.) If desired, you can customize the protocol, such as step duration or number of mixes, right on the screen.
The best way to understand the simplicity of KingFisher sample preparation is to see it in action. We recommend that you schedule a live demo in your lab with your own samples, or a virtual demo we can customize for your lab. To get a flavor of what you’ll experience, watch our demo video.
Live or virtual demos, as illustrated in this video, demonstrate the simplicity of KingFisher sample preparation. (27:28)
After seeing a KingFisher demo in their lab, most customers purchase the instrument for a particular application. But once they install it in their lab and learn out how truly easy it is, they often find other uses for it and it becomes a true laboratory workhorse.
Ct values for contrived SARS-CoV-2–positive nasopharyngeal swab samples with different sample preparation workflows. Nasopharyngeal swab specimens were spiked with 250 copies/mL (left) or 750 copies/mL (right) of SARS-CoV-2 viral RNA (1x or 3x the limit of detection, respectively), and RNA was extracted from 200 μL or 400 μL of the spiked sample in triplicate using the MagMAX Viral/Pathogen Kit with (+) or without (–) Wash 3. After elution in 50 μL, the samples were assayed by qRT-PCR for four RNA targets (colored symbols) in triplicate. Ct values for each target were highly consistent (symbols cluster closely) and virtually identical between sample volumes and wash conditions. All negative samples had Ct values of “Undetected” (data not shown).