Preventing DNA Contamination: Tackling the Phantom of Heilbronn Legacy with Eppendorf

Failures brought to light by botched police investigation have led to tough new requirements for the manufacturers of DNA analysis products

18 Dec 2017
Abigail Berry
Administrator / Office Personnel

Editorial article

Almost ten years ago, detectives in Germany were forced to admit that a supposed female serial killer they had been hunting for over 15 years never existed. The so-called "Phantom of Heilbronn" had previously been connected by DNA evidence to six murders, as well as dozens of other crime scenes. But in 2009, it emerged that the "Phantom" criminal did not exist, and that the DNA in fact belonged to an innocent employee of the factory where the swabs used for collecting DNA samples were made. The confusion led to the introduction of new international standards for the production of kits and consumables for DNA analysis. In this interview, SelectScience speaks to Kayla Hager and Dr. Nils Gerke, of Eppendorf, to discuss how ISO 18385 is changing the manufacturing process for DNA consumables and the implications it has for forensic science. Dr. Gerke spent several years as a forensic DNA analyst prior to joining Eppendorf. Hager was previously a graduate student studying genetics and development before starting her career at Eppendorf North America.

Dr. Nils Gerke (L), Eppendorf global product manager for sample handling consumables. Kayla Hager (R) is the product manager for consumables in North America.

SS: What challenges do forensic scientists face in the field of forensics and DNA analysis?

The prevention of contamination is a major challenge in forensic DNA analysis. In order to minimize the risk of contamination, forensic laboratories have established internal standards and accreditations. However, since the introduction of ISO 18385, there are more demands on the manufacturers of the products used in forensic DNA laboratories.

SS: What is ISO 18385, and why is it important?

ISO 18385 came into effect at the beginning of 2016 and it defines the demands on manufacturers of products used for the collection, storage, and analysis of biological material during the course of forensic investigations. Only products fulfilling this high standard can be labeled as Forensic DNA Grade according to the standard.

An important reason for the creation of ISO 18385 was the case of the “Phantom of Heilbronn”. For more than a decade, police based their investigations of around 40 different criminal cases in Germany, Austria and France on the involvement of one person with a particular DNA profile. In the end, it turned out that the cotton swabs used in the securing of evidence had been contaminated with this very DNA during production. This case was an important cause for the international forensic community to set up the ISO 18385.

The two main objectives of the ISO 18385 are:

Further minimizing the risk of contamination in forensic DNA analysis

  • Manufacturers shall help to avoid human DNA contaminations by implementing appropriate measures in the production process
  • Ensuring and verifying that the final lab products are free of human DNA
  • In case of a contamination in a forensic laboratory:

  • A manufacturer shall help to clear up contaminations by offering a possibility to check a contamination profile against the manufacturer‘s DNA employee exclusion database
  • SS: Describe the steps Eppendorf utilizes to minimize the risk of human DNA contamination

    Eppendorf upholds high-quality standards throughout the manufacturing process of the Forensic DNA Grade product line, ensuring the strict demands of the ISO 18385 standard are met. The risks of contamination during the manufacturing process are minimized by utilizing a high degree of automation, minimalizing direct contact between staff and product whilst monitoring for human DNA in the production environment.

    Tests are performed to verify the certified purity parameters and are carried out at different points of the production process by an independent external testing laboratory. This laboratory is accredited in accordance with ISO 17025 – the same accreditation standard which is applied to forensic laboratories.

    Special attention has also been directed towards the packaging of Eppendorf Forensic DNA Grade products. All Forensic DNA Grade consumables have clearly marked labels on the folding box and also on each individual product bag to ensure unambiguous identification during use. The products are either packaged individually, in process-oriented numbers of items per bag, or in re-sealable bags allowing contamination-free storage. In addition, according to the requirements of the ISO 18385, production-relevant employees are included in a DNA exclusion database, so cases like the “Phantom of Heilbronn” can be avoided.

    SS: What key forensic products does Eppendorf offer for this protocol?

    The Forensic DNA Grade product line encompasses consumables for DNA extraction, sample processing and PCR setup, as well as sample storage. These include:

    SS: What is the future of forensic science, specifically in relation to DNA and contamination?

    The ongoing implementation of next-generation sequencing (NGS) into forensics allows researchers to get more information from forensic samples. By using the targeted NGS approach it is not only possible to get more information, but also specific, forensically relevant information in just a single, targeted run. Alongside this development, the demands on consumables used in forensics with regards to performance and purity will remain on a very high, if not increasing level. Thus, it can be expected that the usage of Forensic DNA Grade consumables according to ISO 18385 will become further established in forensic laboratories worldwide.

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    twin.tec PCR Plate 96 and 384

    Eppendorf

    Eppendorf® twin.tec PCR plates give you the advantages that come from combining two materials: the wells are manufactured from virgin polypropylene, the most successful material used for PCR tubes; polycarbonate, an especially rigid material, forms the plate deck and the skirt. This one-piece design is achieved by an innovative injection moulding process, twin-shot technology. Polycarbonate enhances the mechanical stability and facilitates handling for the twin.tec PCR plates. Its enhanced stiffness guarantees improved plane parallelism in the plate deck before, during and after PCR, thus ensuring a more secure seal. Minimum sagging of the skirt makes the plate the ideal choice for use in automation and in conjunction with robots. All these features are nevertheless of equal use to molecular biologists working in manual operation, because the visible outcome is a batch of more reliable PCR results. Polypropylene gives the wells an inert, non-wetting surface that prevents DNA, RNA and enzymes from binding and improves recovery even when you are working with the most minute volumes. Compared to the common thin-walled tubes, Eppendorf twin.tec PCR plates offer a further 20% reduction in wall thickness. This provides an ideal snug fit for the well against the thermal block and ensures virtually undisturbed heat transfer from the block to the sample. Raised well rims optimize sealing effectivity and reduce the risk of cross contamination. All commonly used sealing methods can be applied: cap strips, adhesive films and foils - and for best protection against evaporation, heat sealing.Eppendorf's certified purity standard - "PCR clean" - guarantees that the product is free of human DNA, DNase, RNase and PCR inhibitors.

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    Safe-Lock micro test tubes

    Eppendorf

    The patented hinged lid on these tubes eliminates any danger of leaks. Safe-Lock micro test tubes guarantee absolute safety during work with expensive or toxic samples, radioactive substances or DNA. The 0.5 ml, 1.5 ml and 2.0 ml test tubes have a graduation scale and a writing surface. The 0.5 ml Safe-Lock micro test tube is ideal for all PCR experiments.Available as Standard Qualitry, PCR Clean and Biopure, and in assorted colors.

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    Combitips®

    Eppendorf

    Eppendorf Combitips® and Combitips® plus form a reliable and precise system for positive-displacement dispensing in combination with the Multipette 4780. Combitips® are also available in Eppendorf Biopur quality, which stands for utmost purity and reliability.

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    PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Sample ManagementSample management systems include sample storage devices such as freezers and plate storers, sample environment enclosures and sample organization, retrieval and sorter systems. Useful system features include high-throughput, automation, robotic arms, automated liquid handling and associated database systems. Accessories in sample management include barcode scanners, heat sealers and tubes.DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA SequencingDNA sequencing, such as sanger sequencing, is a biological technique that determines the precise order of nucleotide bases in a fragment or template of DNA. DNA sequencers and genetic analyzers are based on capillary electrophoresis, where labeled DNA fragments are electrophoretically separated by size as they migrate through a polymer. Find the best DNA sequencing products, including DNA sequencing kits, genomic libraries and genetic identity kits in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Robotic InstrumentsRobotic instruments can be used for high-throughput automation of many lab processes. Such processes use instruments for assays like cell based assays and ELISA, for sample preparation like shakers, centrifuges and incubators, and for analysis such as sequencing and western blot analyzers. Useful features of robotic instruments include speed, reproducibility, barcode readers, software and automation.Robotic WorkstationsRobotic workstations are automated platforms enabling the user to perform high-throughput, multi-application processes, from sample preparation tasks such as PCR set-up, DNA purification and NGS library preparation, to integrated sample prep and analysis workstations. Automation makes processes consistent, fast, precise and fully walk-away. Find the best robotic workstations in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Automated Liquid HandlingAutomated liquid handling equipment is used to mix, dilute and dispense allotted volumes of liquid into microplates and other vessels automatically. The robotic, liquid handling arms can dispense from single channel to 3456 multichannel as well as operate nanoliter dispensing, enabling high throughput of samples. Find the best automated liquid handlers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Safety EquipmentLaboratory safety equipment is essential to the effective operation of all laboratories and is required to minimize the risk of harm during experiments. Safety equipment includes radiation safety, radioactive / hazardous waste disposal, alarms and personal safety.Evidence Collection and AnalysisEvidence Collection and Analysis is the retrieval and subsequent investigation into criminal evidence obtained from a crime scene. Forensic genetic instruments, kits and reagents enable the analysis of evidence collected  at the crime scene. Reagents and kits for presumptive analysis for body fluids such as blood, saliva and semen in addition to drugs and narcotics test kits are used for CSI. Fingerprinting equipment, forensic light sources, casting kits and equipment for collecting recoverable materials can be found in the directory.AutomationAutomation in laboratories and manufacturing processes enhances efficiency, precision, and scalability by reducing the need for manual intervention. It plays a critical role in improving productivity, minimizing human error, and accelerating workflows in fields like diagnostics, drug development, and industrial testing. Automation technologies include robotic systems, automated liquid handlers, and process control systems that streamline complex tasks and ensure consistent, reproducible results. Explore our peer-reviewed product directory to discover the best automation solutions, compare options, read user reviews, and get prices directly from manufacturers.Process DevelopmentProcess development aims to optimize the performance of manufacturing systems. This involves the assessment of quality and efficiency of both processes and products. This is particularly important in the scale-up of chemical processes.ISO CertificationForensic InvestigationForensic investigation is a diverse collection of scientific techniques and methods for the identification and characterization of chemicals relating to crime scene investigations.Process Monitoring