For additional performance data for specific kits and reagents, follow their Cat. No. links in the figure captions. For additional application data, download the application notes listed in this section and on the Resources page.
Sequencing consistency of FFPE cancer samples
DNA and RNA were extracted from matched core needle biopsies and fine-needle aspirate samples using the MagMAX FFPE DNA/RNA Ultra Kit (Cat. No. A31881) on the KingFisher Duo prime. Libraries were prepared and samples were templated and sequenced. Sequencing metrics such as (A) mean read length and (B) DNA mapping and uniformity were consistent, demonstrating functional recovery of nucleic acids from different specimen types.
miRNA and protein analysis from liquid biopsy samples
miRNA biomarker screening for prostate cancer
Total RNA was isolated from the serum of 10 individuals with prostate cancer and 10 healthy individuals using the MagMAX mirVana Total RNA Isolation Kit (Cat. No. A27828) on the KingFisher Flex. Expression levels of 19 miRNAs were measured by qRT-PCR using the TaqMan Advanced miRNA cDNA Synthesis Kit (Cat. No. A28007) and TaqMan Advanced miRNA Assays (Cat. No. A25576). The y-axis displays ΔCt values for each miRNA, normalized to reference mRNA. Large differences between healthy male samples (blue) and prostate cancer samples (green) identify potential cancer biomarkers.
Exosome pull-down
CD9+ exosomes from SW480 colon cancer cells were isolated on the KingFisher Flex by targeting CD9 followed by staining and analyzed by flow cytometry. CD9– exosomes from Jurkat cells were included as controls. Histograms show a distinct CD9+ population (red) among the SW480 cells (B) compared to Jurkat controls (C). The scatter plot and gating profile is also shown (A).
DNA and RNA isolation from hematologic samples
Recovery and purity of DNA from whole blood
A KingFisher Duo Prime was used to isolate DNA from two donors’ whole blood using the MagMAX Multi-Sample Ultra 2.0 Kit (Cat. No. A36570). Samples >400 µL were processed using the large-volume protocol in a 24-well plate, while samples 50–400 µL were processed using the small volume protocol in a 96-well plate. (No sample volume normalization was needed, even when 50 µL and 400 µL samples were run at the same time on the same plate.) Run time was 45 minutes. (Top) Recovery volumes were linear across both small and large volume inputs. (Bottom) A260/A280 and A260/A230 ratios demonstrated high purity across sample volumes.
Sequential DNA and RNA isolation from whole blood and bone marrow
Sequential isolation performance data for DNA using bone marrow and blood sample types
DNA averaged across multiple samples across various sample volume input. DNA measured on Invitrogen Qubit Flex Fluorometer with Invitrogen Qubit 1X dsDNA Broad Range Assay. Please note that DNA yields are variable from donor to donor and dependent on sample quality, age and input.
RNA averaged across multiple samples across various sample volume input. RNA measured on Invitrogen Qubit Flex Fluorometer with Invitrogen Qubit RNA High Sensitivity (HS) Assay. Please note that RNA yields are variable from donor to donor and dependent on sample quality, age and input.
Nucleic acid isolation for bacterial and viral detection
Microbiota identification
Total nucleic acid from fecal, urine, and saliva samples from two donors was isolated using the MagMAX Microbiome Ultra Nucleic Acid Isolation Kit (Cat. Nos. A42357 and A42358) on the KingFisher Flex, submitted for shotgun metagenomic sequencing, and compared with known microbial profiles. The figure shows abundance heat maps for selected microbiota for all samples. Results confirmed that each body habitat harbored different dominant signature taxa, with only a few shared species, mostly between urine and saliva. In each community, there was substantial overlap between donors but also marked individual differences.
Veterinary pathogen detection
For a study at the University of California Davis School of Veterinary Medicine, whole blood, feces, and nasal secretions were obtained from canine, feline, and equine species. Total nucleic acids were extracted on the KingFisher Flex and a vacuum-based automated system and amplified with qPCR. Quantification cycle (Cq) values were comparable for the two methods and the total number of pathogens detected was similar. However, the extraction time with the KingFisher Flex was three times faster.
SARS-CoV-2 testing
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 (Cat. No. A42352) 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, validating the streamlined lower-volume protocols. All negative samples had Ct values of “Undetected” (data not shown).
Automated preparation for protein analysis
Immunoprecipitation
Jurkat cells expressing CD81 were lysed and incubated with Dynabeads Protein G (Cat. No. 10004D) coated with an anti-CD81 antibody. CD81 was isolated from the cell lysate in triplicate using an optimized walkaway IP protocol on the KingFisher Duo Prime and Flex instruments and compared with the manual protocol. Electrophoresis and western blotting confirmed good performance on the two instruments, equivalent to the results obtained with the manual protocol.
Peptide mapping
The drug infliximab was digested using the SMART Digest Trypsin Kit on the KingFisher Duo Prime. Analysis of the UV peptide map by LC-MS confirmed complete sequence coverage of 100% for both the light and heavy chain of the antibody. In other data, the automated protocol was found to be reproducible and easy to use, even by those who had no prior experience with protein digestion.
For Research Use Only. Not for use in diagnostic procedures.