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Extraction – Vacuum based

Extraction Vacuum Based_1

Extraction technologies based on binding matrices integrated in spin columns or binding plates are state of the art technology used already for a long time in molecular laboratories around the world.

However, the respective kits are tedious when manually performed and need additional equipment like centrifuges or vacuum devices.

Hamilton provides flexible workstations equipped with a fully automated and integrated vacuum station for the extraction or purification of nucleic acids from a great variety of sample sources.

DNA extraction from grapevine

Conventional DNA extraction methods for PCR-quality DNA from plants are tedious. When a large number of samples need to be analyzed, e.g. in breeding programs for improving quality, performance or robustness of cultured plants, a rapid and simple procedure is required.

DNA extraction from plants is a very demanding procedure. Numerous secondary metabolites such as polyphenolics, polysaccharides or acidic components can contaminate the sample and require several time-consuming processing steps. Especially for reproducible PCR results, a protocol yielding high quality DNA in a sufficient quantity is required. Here, we present an automated and efficient method for genomic DNA extraction from grapevine implemented on the Microlab STARlet liquid handling robot using the NucleoSpin® 8 Plant kit from MACHEREY-NAGEL and assess the quality of the extracted DNA.

Features and Benefits

  • MACHEREY-NAGEL NucleoSpin® 8 Plant kit used for the extraction of genomic DNA from plant tissue based on vacuum filtration
  • 48 cleared sample lysates can be processed in less than one hour without user intervention
  • Fully automated process from clarified samples to eluted DNA
  • Adjust laboratory throughput: flexibly load samples to match experimental workflow
  • Extraction yields high quality DNA suitable for PCR and other downstream applications
  • Other downstream applications can be implemented on the same platform taking advantage of Hamilton’s highly flexible and modular pipetting workstation

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Automated genomic DNA Isolation from whole Blood on the Genomic STARlet

Extraction of genomic DNA out of blood samples is a time-consuming and often limiting factor for genotyping applications. Automated processing of this sample material is highly desired, but in addition, it requires thorough process control for liquid handling of this difficult biological source.
The analysis of genomic DNA from whole blood is widely used for genotyping, HLA typing, and for other diagnostic procedures. It is relatively simple to draw the blood samples. The processing, however, offers some challenges due to the possible clotting of the blood. Therefore a reliable system and process control are crucial for isolation of genomic DNA from whole blood samples, which are stabilized using EDTA, citrate or heparin. Hamilton and MACHEREY-NAGEL have developed the Genomic STARlet to isolate genomic DNA from blood samples using the NucleoSpin® technology. This application note describes automated isolation of genomic DNA from blood and compares the results with a manual isolation process. It further shows how Hamilton’s monitored air displacement pipetting principle allows for detection of erroneous pipetting steps and helps to increase process stability.

Features and Benefits

  • Address time-consuming sample preparation for genotyping
  • Gain control over DNA isolation process
  • Make DNA isolation from whole blood reliable
  • 48 samples with the 8-well strips or 96 samples with the 96-well binding plate can be processed in less than 80 minutes
  • Designed for optimal reliability, throughput, yield and quality for the extraction of nucleic acids from various blood samples such as fresh and frozen blood from humans or animals
  • Automated and manually isolated genomic DNA out of human fresh blood gave similar results in yields and homogenous qualities of DNA.
  • Hamilton’s pressure based total aspiration monitoring detects clogging of columns and exclusion of clogged columns from further processing by the software increases process stability
  • Better standardization and consistent DNA quality for downstream applications such as genotyping and HLA typing

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Automated genomic DNA Isolation from Mouse Tails on the Genomic STARlet

Large-scale studies of genetic variations or transgene animal identification require parallel processing of a large number of biological samples such as tissue. Reliable handling of these samples and a suitable process control are essential for automating genotyping applications.
Genotyping methods like restriction-, qPCR-, and SNP-analysis are widely used in research and diagnostics of diseases. DNA of high quality is required for a reliable analysis. Hamilton and MACHEREY-NAGEL have developed the Genomic STARlet to isolate DNA from different samples such as cells and tissues using the NucleoSpin® technology. This application note describes the automated isolation of DNA from mouse tails and compares the results with a manual isolation process. Hamilton and MACHEREY-NAGEL have developed and verified a method for fully automated genomic DNA isolation from tissue on the Genomic STARlet with maximum reliability, throughput, yield and quality.

Features and Benefits

  • Increase throughput for genotyping methods
  • Avoid contamination
  • Ensure consistency in tissue processing
  • Genomic STARlet processed 48 samples with the 8-well strips or 96 samples with the 96-well binding plates in less than 80 minutes
  • No carryover or cross-contamination as demonstrated by qPCR
  • Hamilton and MACHEREY-NAGEL have designed the Genomic STARlet to allow optimal reliability, throughput, yield and quality for the extraction of nucleic acids from various samples, e.g. mouse tail end clippings.
  • Automated and manual isolation resulted in similar yields and qualities of DNA.
  • The use of the MN Wash Plate during vacuum processing effectively prevents ethanol contamination derived from ethanolic wash buffers.
  • Automated isolation of DNA from tissues on the Genomic STARlet guarantees increased standardization for downstream genotyping applications

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Automated Plasmid Purification on the Genomic STARlet

Isolation of plasmid DNA is a bottleneck in many research or diagnostic laboratories. Fast and efficient automation of plasmid purification is required in order to replace labor intensive manual work. High purity and yields must be achieved in a reproducible procedure.

Flexible and efficient workstations for isolation of DNA have become more important in the past few years in many areas of life sciences as sample preparation is a time-consuming step in genetic analysis. As a solution to this problem, Hamilton and MACHEREY-NAGEL have designed the Genomic STARlet for different applications, e.g. plasmid DNA purification. The flexible purification system is based on the liquid handling robot Microlab STAR and suitable for use of the silica membrane based NucleoSpin® 8/96 Plasmid Purification Kits from MACHEREY-NAGEL.

Features and Benefits

  • Save time in sample preparation for genetic analysis
  • Include all steps required for DNA purification process
  • Work reliably even when low DNA quantities are expected
  • Validated for the isolation of plasmids from bacteria
  • Plasmids can be directly used in downstream applications such as sequencing
  • Yield and purity from automated and manual isolation processes comparable
  • Increased standardization by reducing errors
  • Simultaneous processing of flexible numbers of samples
  • Additional kits can be processed on the same system

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Plasmid DNA Purification using the Promega Wizard SV 96 Plasmid Purification System

Plasmid DNA is useful in a wide range of molecular biology applications.  However, purification of plasmid DNA from pelleted bacterial culture can be time-consuming when done manually.  Automation of this process can deliver significant increases in throughput as well as higher levels of precision and accuracy.  We have developed an automated method that enables processing of multiple 96-well plates per day using the Promega Wizard SV 96 Plasmid Purification System on the Hamilton MICROLAB STAR workstation.  Here we provide an example of high-copy plasmid DNA purification with this easy-to-follow automated protocol.  High-purity plasmid DNA is isolated consistently from bacterial culture samples with no detectable cross-contamination, and can be used directly in downstream sequencing and restriction enzyme digestion.

Features and Benefits

  • Successful isolation and recovery of high-quality plasmid DNA directly from pelleted bacterial culture. Coupled with Hamilton’s
  • Promega Wizard SV 96 Plasmid DNA Purification System represents a high-throughput, easy-to-use approach for reliable recovery of purified DNA fragments
  • DNA fragments are suitable for many downstream applications
  • No cross-contamination observed

Automated Total RNA Isolation from Cells and Tissues on the Genomic STARlet

Genome-wide transcriptional profiling is essential for exploring a biological mechanism or predicting disease. It requires isolation of high quality RNA from a large number of biological samples such as cultured cells or tissue which represent different conditions under investigation.

High purity and integrity of RNA are crucial for reliable gene expression studies. Hamilton and MACHEREY-NAGEL have developed the Genomic STARlet to isolate nucleic acids from different samples such as cells and tissues using the NucleoSpin® technology. This application note describes the automated isolation of RNA from HeLa cells as well as from mouse liver and compares the results with a manual isolation process.

Features and Benefits

  • Isolate RNA with high integrity
  • Address large sample numbers for gene expression studies
  • Ensure reliability for processing of cells and tissue
  • Hamilton and MACHEREY-NAGEL have designed the Genomic STARlet to allow optimal reliability, throughput, yield and quality for the extraction of nucleic acids from various samples, as demonstrated for total RNA from HeLa cells and mouse liver tissues
  • Automated and manual isolation resulted in similar yield and quality of RNA.
  • Isolate high quality RNA for direct use in further downstream applications as rt-qPCR analysis.
  • Automated isolation of RNA on the Genomic STARlet guarantees increased standardization for downstream gene expression studies

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Sample preparation for viral diagnostics in veterinary applications - BVD Diagnostics

Detection of viral pathogens for animals in veterinary laboratories involve serological as well as DNA based diagnostic tests. Large numbers of potentially infectious samples must be processed periodically in a short time. It is desired to automate a number of different tests on the same platform.

Surveillance testing for avian influenza and bovine virus diarrhea (BVD) is becoming an increasing challenge for veterinarian laboratories. Routine surveillance already reaches 3 million tests every year in European countries but reaches seasonal peaks that are particular challenging. In addition to serological testing, nucleic acid based testing that investigates presence of viral RNA or DNA in samples is becoming a powerful, economical and fast alternative. Since viral RNA is present long before viral protein can be detected, this method also closes the diagnostic gap between infection and outbreak of the disease. The Microlab STAR liquid handling platform is used for automated isolation of viral RNA using the NucleoSpin® 96 Virus kit from MACHEREY-NAGEL. Isolated viral RNA is subsequently used to detect BVD virus (BVDV) in serum using real-time qPCR.

Features and Benefits

  • Ensure process safety for working with infectious samples
  • Provide a high level of precision and accuracy
  • Use one system for various diagnostic applications
  • MACHEREY-NAGEL’s NucleoSpin® 96 Virus kit for isolation of viral RNA or viral DNA in combination with the Microlab STAR, offer a fully automated RNA preparation system for ultimate process safety
  • Monitored Air Displacement in combination with Hamilton’s CO-RE technology offers contamination free, fully automated BVD testing
  • The NucleoSpin® 96 Virus kit has also been successfully used to isolate RNA from other samples, for example avian influenza virus RNA from infected birds or blue tongue virus from cattle blood making it a complete solution for the veterinarian laboratory

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