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Microscale thermophoresis analysis

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An emerging and sensitive method for studying molecular interactions is microscale thermophoresis (MST). The technique is based on the motion of molecules along local temperature gradients, measured by fluorescence changes [ 21, 22 ]. A spatial temperature difference (Δ T) leads to a mass flow of molecules in the temperature-elevated region Microscale Thermophoresis SOME STARTUP LIKE IT HOT. What is Microscale Thermophoresis? Technique for the analysis of molecule binding interactions uses the thermophoretic effect on a microscale allows experiments under almost natural conditions (blood serum) is able to determine the dissociation constant of binding events by measuring under different ligand concentrations (titration) 2. Microscale thermophoresis (MST) can be widely used for determination of binding affinity without purification of the target protein from cell lysates. The protocol involves overexpression of the GFP-fused protein, cell lysis in non-denaturing conditions, and detection of MST signal in the presence of varying concentrations of the ligand

Microscale thermophoresis - Wikipedi

Microscale Thermophoresis (MST) is a powerful new technology, and easy to handle. It detects changes in the hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice While many biophysical methods may be used to obtain KD, the focus of this report is a relatively new method called microscale thermophoresis (MST). In an MST experiment, a capillary tube filled with a solution containing a dye-labeled solute is illuminated with an infrared laser, rapidly creating a temperature gradient

MicroScale Thermophoresis: Interaction analysis and beyond

Establishment of a novel microscale thermophoresis ligand-binding assay for characterization of SLC solute carriers using oligopeptide transporter PepT1 (SLC15 family) as a model system. Clémençon B(1), Lüscher BP(2), Hediger MA(3). Author information: (1)Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland The emergence of microscale thermophoresis (MST) as a technique for determining the dissociation constants for bimolecular interactions has enabled these quantities to be measured in systems that were previously difficult or impracticable. However, most models for analyses of these data featured the assumption of a simple 1:1 binding interaction

Microscale thermophoresis (MST) is a biophysical technique that has seen increasing application in the study of biomolecular interactions thanks to its solution-based nature, its rapid application, modest sample demand, and the sensitivity of the thermophoresis effect to binding events MicroScale Thermophoresis (MST) is a powerful technique to quantify biomolecular interactions. It is based on thermophoresis, the directed movement of molecules in a temperature gradient, which..

Microscale Thermophoresis. Messung der Thermophorese eines fluoreszenzmarkierten Biomoleküls. Die normierte Fluoreszenz im erwärmten Laser-Spot ist gegen die Zeit aufgetragen. Der IR-Laser wird zum Zeitpunkt t=5s eingeschaltet und induziert eine messbare Änderung der Fluoreszenz field of high-throughput protein stability screenings based on MicroScale Thermophoresis (MST) were performed in close collaboration with NanoTemper Technologies GmbH, Munich. First and foremost, I want to express my deepest gratefulness to my doctoral advisor and mentor Prof. Dr. Gerhard Winter for the excellent guidance and continuous support throughout my time at the university and during. Foley et al. Analysis of conglutin seed storage proteins across lupin species using transcriptomic, protein and comparative genomic approaches. BMC Plant Biology (2015) 15(106): 1-12. Gupta, Duhr & Baaske. MicroScale Thermophoresis (MST). Encyclopedia of Biophysics. 2018: 1-5. Siller-Matula et al. Thrombin as a multi-functional enzyme. Focus on. Microscale thermophoresis (MST) allows for quantitative analysis of protein interactions in free solution and with low sample consumption. The technique is based on thermophoresis, the directed motion of molecules in temperature gradients. Thermophoresis is highly sensitive to all types of binding-induced changes of molecular properties, be it in size, charge, hydration shell or conformation.

A recently developed technology, Microscale Thermophoresis (MST), uses this temperature field to perform biomolecular interaction studies. Thermophoresis, the motion of molecules in temperature fields, is very sensitive to changes in size, charge, and solvation shell of a molecule and thus suited for bioanalytics Microscale Thermophoresis Assay Services MST assays measure the motion of molecules along microscopic temperature gradients that changes upon ligand binding. MST assays measure the ligand-target interactions directly in solution without the need for immobilization to a surface (immobilization-free technology) One way to bypass these restrictions is to use the relatively new technique of microscale thermophoresis (MST). MST is fast and cost effective using small amounts of sample to obtain a saturation curve for a given binding event. There currently are two types of MST systems available. One type of MST requires labeling with a fluorophore in the blue or red spectrum. The second system relies on. In this video, I am trying to explain MST technology in simple terms with analogies! I hope this helps This manual is your guide for using the Monolith NT.115 and doing Microscale Thermophoresis (MST) measurements. It instructs first-time users on how to use the instrument, and serves as a reference for experienced users. Before using the Monolith NT.115 instrument, please read this instruction manua

MO.Affinity Analysis 3 Make sure your analysis is consistent across various data sets and that you're identifying any insights across replicate measurements. MO.Affinity Analysis 3 is the ideal complement to MO.Control 2 — merge and group your data sets for comparison purposes and then easily report results with presentation-worthy data and publication-ready figures Microscale thermophoresis (MST) allows for quant. anal. of protein interactions in free soln. and with low sample consumption. The technique is based on thermophoresis, the directed motion of mols. in temp. gradients. Thermophoresis is highly sensitive to all types of binding-induced changes of mol. properties, be it in size, charge, hydration shell or conformation. In an all-optical approach. High-quality biophysical characterization tools to help researchers study challenging molecular interactions, protein stability, and protein characterization Microscale thermophoresis (MST) is a technology for the biophysical analysis of interactions between biomolecules. Microscale thermophoresis is based on the detection of a temperature-induced change in fluorescence of a target as a function of the concentration of a non-fluorescent ligand. The obse

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MicroScale Thermophoresis • MST • Binding affinity

A Brief Comparison of Microscale Thermophoresis (MST) and Isothermal Titration Calorimetry (ITC) Posted on April 5, T. André, R. Wanner, H. M. Roth, S. Duhr, P. Baaske and D. Breitsprecher (2014). MicroScale Thermophoresis: Interaction analysis and beyond. Journal of Molecular Structure 1077: 101-113. Leavitt, S. and E. Freire (2001). Direct measurement of protein binding. Herein, we present a hybridization‐based approach that uses microscale thermophoresis (MST) as a very fast and highly precise readout to quantify, for example, single tRNA species with a turnaround time of about one hour. We developed MST to quantify the effect of tRNA toxins and of heat stress and RNA modification on single tRNA species The thermophoresis signal is plotted against the ligand concentration to obtain a dose-response curve, from which the binding affinity can be deduced. The dynamic affinity range covers the pM to mM range, which is comparable to SPR measurements. MST requires a small amount of sample and the technique is relatively easy to use MicroScale Thermophoresis (MST) is based on the directed movement of molecules in a temperature gradient which strongly depends on a variety of molecular properties such as size, charge, hydratation shell or conformation thus being a powerfull technique to quantify biomolecular interactions

Microscale thermophoresis as a powerful tool for screening

Bei einem neueren Verfahren, der optisch erzeugten Thermophorese (englisch: Microscale Thermophoresis), wird mit Hilfe eines Infrarot- Lasers in einer flüssigkeitsgefüllten Glaskapillare ein definierter mikroskopischer Temperaturgradient erzeugt The method utilizes the autofluorescence of synthetic melanin in microscale thermophoresis (MST), and thereby avoids ligand quantitation with expensive mass spectrometry. Dissociation binding constants (K d) obtained by MST were compared to the binding affinities from traditional melanin binding assays

Microscale thermophoresis (MST) is an emerging technique that has been broadly applied to investigate biomolecular interaction of a variety of drug targets. MST detects the directed movement of fluorescent molecules in microscopic tempera-ture gradients in microliter-volume capillaries to quantify interaction affinities4,5 (Suppl. Fig. 1). Each molecule has dis-tinct thermophoretic properties. This manual is your guide for using the Monolith NT.115 and doing MicroScale Thermophoresis (MST) measurements. It instructs first-time users on how to use the instrument, and serves as a reference for experienced users. Before using the Monolith NT.115 instrument, please read this instruction manual carefully, an A recently developed technology, Microscale Thermophoresis (MST), uses this temperature field to perform biomolecular interaction studies. Thermophoresis, the motion of molecules in temperature fields, is very sensitive to changes in size, charge, and solvation shell of a molecul Label-Free Microscale Thermophoresis Discriminates Sites and Affinity of Protein-Ligand Binding** Susanne A. I. Seidel, Christoph J. Wienken, Sandra Geissler, Moran Jerabek-Willemsen, Stefan Duhr, Alwin Reiter, Dirk Trauner, Dieter Braun, and Philipp Baaske* The analysis of protein binding to small molecules, nucleic acids, and ions not only gives fundamental insights into cellular processes.

Protein Purification-free Binding Analysis by MicroScale

  1. Quantitative analysis of protease recognition by inhibitors in plasma using microscale thermophoresis Skip to main content Thank you for visiting nature.com
  2. microscale thermophoresis 4.1.2 Chemical shift perturbation analysis (CSP) 78 4.1.3 Affinity of binding 81 4.1.4 Paramagnetic relaxation enhancement (PRE) analysis 81 4.1.5 Dynamic equilibrium of VpUcyt conformations and binding to CD4mut 85 4.1.6 Biological context 87 4.2 Microscale Thermophoresis (MST) 90 4.2.1 Rationale for employing nanodiscs in microscale thermophoresis (MST) 90 4.2.2.
  3. The analysis of binding interactions between small molecules and biopolymers is important for understanding bio-logical processes. While fluorescence correlation spectroscopy (FcS) requires fluorescence labeling on the small mole-cule, which often interferes with binding, in microscale thermophoresis (MST) the label can be placed on the biopolymer

Microscale Thermophoresis (MST) MST is performed using thin capillaries in free solution, which is comparable to ITC measurements. When performing an MST experiment, a microscopic temperature gradient is induced by an infrared laser, and the directed movement of molecules is detected by intrinsic fluorescence or in most cases, fluorescent labels of one interactant and quantified gleichen biophysikalischen Analysemethode der Mikrothermophorese (MST). Teil I konzentriert sich auf die präbiotische Replikation von DNA. Verschiedene Ansätze zur Rep- likation von RNA und DNA unter präbiotischen Bedingungen werden in der Einführung diskutiert Microscale thermophoresis (MST) is a technology for biomolecules interaction analysis. MST is a rapid and precise method to quantify biomolecular interactions in solution at microliter scale. Microscale thermophoresis is the directed movement of particles in a microscopic temperature gradient. Any change of the hydration shell of biomolecules due to changes in their structure/conformation. Article Title: Using Microscale Thermophoresis to Characterize Hits from High-Throughput Screening: Data analysis was performed with the MO Affinity Analysis software (Nanotemper), and the data was fit using the specific binding with Hill model in GraphPad.. TDP-43 proteins (0.75 nM) were mixed with 10 μ g/mL of anti-His acceptor beads (PerkinElmer) and RNA sequences (biotinylated-UG6. Protein interactions inside the human body are expected to differ from the situation in vitro. This is crucial when investigating protein functions or developing new drugs. In this study, we present a sample-efficient, free-solution method, termed microscale thermophoresis, that is capable of analysing interactions of proteins or small molecules in biological liquids such as blood serum or.

On the acquisition and analysis of microscale thermophoresis data. Analytical Biochemistry 2016, 496, 79-93. DOI: 10.1016/j.ab.2015.12.013. Hermann Wätzig, Imke Oltmann-Norden, Franziska Steinicke, Hassan A. Alhazmi, Markus Nachbar, Deia Abd El-Hady, Hassan M. Albishri, Knut Baumann, Thomas Exner, Frank M. Böckler, Sami El Deeb. Data quality in drug discovery: the role of analytical. MicroScale Thermophoresis (MST) is an exceptionally powerful technique to quantify biomolecular interactions. It is based on thermophoresis, the directed movement of molecules in a temperature gradient, which strongly depends on a variety of molecular properties such as size, charge, hydration shell and conformation. Thus, this technique is highly sensitive to virtually any change in molecular.

The analysis software is used to calculate the dissociation coefficient. Microscale Thermophoresis can monitor the binding of single ions (40Da) or small molecules (300Da) to a target as well as the binding of ribosomes (2.5MDa). The capillary format is easy to handle and offers maximum flexibility in the experiment scale. The capillaries come with different surface coatings to stabilize even. MicroScale thermophoresis (MST) is a new method that enables the quantitative analysis of molecular interactions in solution at the microliter scale and high sensitivity. The technique is based on the effect that molecules move in temperature gradients, a physical effect called thermophoresis. The thermophoresis of molecules depends o Innovative Technologie zur Analyse von Biomolekülen: Microscale Thermophoresis Mittwoch, 20. Juli 2011, 15:00 bis ca. 19:00 Uhr Veranstaltungsort: NanoTemper Technologies GmbH, Flößergasse 4, 81369 München Download Programmvorschau hier als PDF-Date Quantitative analysis of such interactions in vivo is essential to study diseases, but have not been forthcoming, as most methods cannot be directly applied in a complex biological environment. Here, we report a quantitative microscale thermophoresis assay capable of deciphering functional deviations from in vitro inhibition data by combining concentration and affinity measurements. We.

On the acquisition and analysis of microscale

interaction analysis by MicroScale Thermophoresis Tanja Bartoschik1, Stefanie Galinec1, Christian Kleusch1, Katarzyna Walkiewicz1, Dennis Breitsprecher1, Sebastian Weigert2, Yves A. Muller 2, Changjiang You 3, Jacob Piehler3, Thomas Vercruysse4, Dirk Daelemans 4 & Nuska Tschammer1 MicroScale Thermophoresis (MST) is a frequently used method for the quantitative characterization of. Analysis of translationally active RNAs : quantification by microscale thermophoresis and modification analysis by LC-MS/MS: Online publishing date: 20-Dec-2016: Language : english: Abstract: RNA, und tRNA im Besonderen, spielen nicht nur in grundlegenden zellulären Prozessen wie der Translation der genetischen Information eine wichtige Rolle, sondern erfüllen auch eine Vielzahl an. Protein Purification-free Method of Binding Affinity. Munich led by Professor Dieter Braun, a member of the Cluster of Excellence Nanosystems Initiative Munich (NIM), and a partner in NanoTemper (an LMU spin-off), have now developed a unique technology called microscale thermophoresis that allows to measure intereactions under close-to-native conditions, thus improving the decision making process in drug development The Protein Interactions Analysis capability enables the study of protein-protein and protein-ligand interactions using various techniques including, analytical ultracentrifugation, to determine the native mass, stoichiometry and shape of proteins and biomolecular complexes in solution, isothermal titration microcalorimetry (ITC) and microscale thermophoresis (MST) to measure the.

Establishment of a novel microscale thermophoresis ligand

New to our lab is the NanoTemper Monolith NT.115pico which uses microscale thermophoresis technology to measure binding affinity between bimolecular interactions from 1pm to mM in complex bioliquids such as cell lysate and serum without immobilization via precise fluorescent dye or fluorescent fusion protein detection. It is also capable of stoichiometry, enthalpy and enzyme kinetics. BioTek. The Complete Monotub Tek Cultivation Walk-through - The Easiest Way to Grow Mushrooms Indoors! - Duration: 36:30. North Spore Recommended for yo Criterion Surface Plasmon Resonance (SPR) Isothermal Titration Calorimetry (ITC) Microscale Thermophoresis (MST) Sample consumption Cost of consumables high low low Capacity/run 1 sample 1 sample 1 - 16 samples Time required for experiment plus analysis hours hours minutes Sample range (Dalton) 10. 2 - 1 MicroScale Thermophoresis: Interaction analysis and beyond. M Jerabek-Willemsen, T André, R Wanner, HM Roth, S Duhr, P Baaske, Journal of Molecular Structure 1077, 101-113, 2014. 344: 2014 : Extreme accumulation of nucleotides in simulated hydrothermal pore systems. P Baaske, FM Weinert, S Duhr, KH Lemke, MJ Russell, D Braun. Proceedings of the National Academy of Sciences 104 (22), 9346.

Using two-site binding models to analyze microscale

Microscale thermophoresis analysis. The binding affinity of TmPiT for different ligands—phosphate, arsenate, sulfate, and PFA—was measured by microscale thermophoresis using a Monolith NT.115 pico instrument (NanoTemper Technologies, Germany). The binding affinity of TmPiT mutants for phosphate was assessed using a Monolith NT.115 instrument (NanoTemper Technologies, Germany). TmPiT (10 nM. MicroScale Thermophoresis (MST) is an immobilization-free technology for measuring biomolecular interactions. The MST instrument detects the motion of fluorescent molecules along a microscopic temperature gradient, which reflects changes in the molecular hydration shell, charge, or size When performing a MicroScale Thermophoresis (MST) experiment, a microscopic temperature gradient is induced by an infrared laser, and the directed movement of molecules is detected and quantified using either covalently attached dyes, fluorescent fusion proteins or intrinsic tryptophan fluorescence Microscale thermophoresis (MST) is a biophysical technique that has seen increasing application in the study of biomolecular interactions thanks to its solution-based nature, its rapid application, modest sample demand, and the sensitivity of the thermophoresis effect to binding events. Here, we describe the use of MST in the study of chromatin interactions, with emphasis on the wide range of.

Die Microscale Thermophoresis (MST) reagiert dabei sensitiv auf Größenänderungen sowie auf Veränderungen von Wasserhülle und Ladung der Moleküle. Wir können die Wechselwirkung von Bindungspartnern in nahezu jeglichem Größenverhältnis analysieren, sagt Braun Detailed knowledge on the binding modes involved is crucial for our understanding of how these protein-protein and protein-DNA interactions shape the functional landscape of chromatin in all kingdoms of life: bacteria, archaea, and eukarya.Microscale thermophoresis (MST) is a biophysical technique that has seen increasing application in the study of biomolecular interactions thanks to its. Microscale thermophoresis: |Microscale thermophoresis (MST)| is a technology for the analysis of |biomolecules|. Microscale World Heritage Encyclopedia, the aggregation of the largest online encyclopedias available, and the most definitive collection ever assembled Microscale thermophoresis revisited One of the less commonly used fragment-finding methods is microscale thermophoresis . This measures the movement of proteins in a temperature gradient; ligand binding changes the movement. When we first described MST in 2012, we noted that the technique seemed relatively low throughput. In a paper recently published in J. Biomol. Screen., Alexey Rak and. MicroScale Thermophoresis binding analysis for the interaction between Spike RBD and Spikeplug. The concentration of NT647-Spike RBD is kept constant at 25 nM, while the ligand concentration varies from 5 µM to 0.15 nM. The serial titrations result in measurable changes in the fluorescence signa

Microscale Thermophoresis Analysis of Chromatin

Beresnev, S. & Chernyak, V. 1995 Thermophoresis of a spherical particle in a rarefied gas: numerical analysis based on the model kinetic equations. Phys. Fluids 7 ( 7 ), 1743 - 1756 The use of microscale thermophoresis in these projects has several advantages over other binding techniques, including low reagent consumption, an unrestricted range of operating concentrations, quick analysis time, and the potential to analyze macromolecules in cell lysates without prior purification MicroScale Thermophoresis technique exploits this effect and is becoming a popular technique for analyzing interactions of biomolecules in solution. When comparing affinities of related compounds, the reliability of the determined thermodynamic parameters often comes into question In the present study, combining the conventional photothermal analysis and the concept of interaction of solvent molecules in interfacial layer used for thermophoresis in liquid, a theory for photophoresis of a hydrophobic particle suspended in liquids is developed. To characterize hydrophobicity of the micro-particle, slip length in Navier's formula is used as an index Microscale Thermophoresis exploits the directed motions of biomolecules and macromolecular complexes in solution in microscopic thermal gradients. It can be used to measure equilibrium binding events. Thermophoretic mobility is dependent on the solvation entropy, charge, size and conformation of the molecules observed

Binding curve generated by the NanoTemper Analysis 1

(PDF) MicroScale Thermophoresis: Interaction analysis and

  1. (2013). Microscale thermophoresis provides insights into mechanism and thermodynamics of ribozyme catalysis. RNA Biology: Vol. 10, No. 12, pp. 1815-1821
  2. Microscale Thermophoresis (MST) is based on a physical principle used for the first time in bioanalytics. It de- tects changes in the hydration shell, charge or size of molecules allowing measurement of a wide range of bio- molecular interactions under close-to-native conditions
  3. Label-free microscale thermophoresis for the study of lactoferrin-drug interaction Mufarreh Asmari1, Ratih Ratih1, Sami El Deeb1 1 Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Beethoven Street 55, 38106 Braunschweig, Germany Correspondence: s.eldeeb@tu-braunschweig.de Abstract In this study, a microscale thermophoretic (MST) method was developed to investigate.
  4. Microscale Thermophoresis Ligand-Binding Analysis Microscale thermophoresis was used to measure the binding interactions between purified receptors and their ligands using a setup similar to that.

Microscale Thermophoresis. The binding of c-di-GMP and related nucleotides to N-terminally FITC-labeled CSP4 (FITC-CSP4-NH 2) was determined on a Monolith NT.115 instrument with standard-treated capillaries (NanoTemper Technologies). Fluorescence changes were recorded using blue channel optics of the instrument for a 30-s period of infrared. • Protein stability analysis/folding • Differential scanning calorimetry • Thermofluor/Stargazer • Circular Dichroism • Protein interactions • Isothermal titration calorimetry • Microscale thermophoresis • Protein biochemistry • Simple Western (automated Western blotting/isoelectric focusing) Microscale Thermophoresis (MST) • Nanotemper Technologies GmbH, founded in 2008.

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Thermophorese - Wikipedi

Materials & methods: We have used MicroScale Thermophoresis analysis and docking calculations to assess on a quantitative basis the binding properties of distinct categories of inhibitors to IDO1 MicroScale Thermophoresis was used to assess the direct interaction between G9a and its inhibitor BIX-01294 and to gain insight into its mode of action (Seidel et al., 2013) 5. With this technology it was possible to detect direct binding interactions between G9a and BIX-01294 Microscale thermophoresis (MST) measures the size, charge and hydration entropy of molecules/substrates at equilibrium. The thermophoretic movement of a fluorescently labeled substrate changes significantly as it is modified by an enzyme. This enzymatic activity can be measured with high time resolution in real time This gel image is representative of three repeated experiments. d Binding experiments were conducted using a microscale thermophoresis (MST) system. The binding affinities of guide RNAs and the effector proteins (SpCas9 and AsCpf1) were measured using Monolith NT.115 (NanoTemper Technologies GmbH) MicroScale Thermophoresis (MST) is a powerful technique to quantify biomolecular interactions. It is based on thermophoresis, the directed movement of molecules in a temperature gradient, which strongly depends on a variety of molecular properties such as size, charge, hydration shell or conformatio

Label-Free Interaction Analysis – WISBMicroscale Thermophoresis (MST) - Integrated StructuralMST Systems for Binding Affinity & Protein InteractionsAssay Development Services - Base Pair Biotechnologies

As an alternative technique, we will use Microscale Thermophoresis (MST) to determine our binding affinity. This is a relatively new technique that employs the phenomenon of thermophoresis, where molecules move through a temperature gradient. This phenomenon is dependent on size, charge, and solvation entropy of the molecule. Therefore, a binding event, such as a transcription factor binding. MicroScale Thermophoresis (MST) measures the motion of molecules along microscopic temperature gradients and detects changes in their hydration shell, charge or size. When performing a MicroScale Thermophoresis (MST) experiment, a microscopic temperature gradient is induced by an infrared laser, and the directed movement of molecules is detected and quantified using either covalently attached. M ST Technology www.nanotemper.de Optical MicroScale Thermophoresis Basic Setup of the Instrument LED excitation & fluorescence detection IR laser induced Temperature Field (BCECF-dye in TRIS-buffer) 23°C dichroic mirror IR laser 20°C objectiv e capilla ry 1K/10 ?m corresponds to 1000K/cm ? ? ? immobilization-free in any buffer 4 ?l of sample per titration point pM - nM concentrations of. DOI: 10.1007/978-1-4939-3197-2_8 Corpus ID: 30784752. Aptamer Binding Studies Using MicroScale Thermophoresis. @article{Breitsprecher2016AptamerBS, title={Aptamer Binding Studies Using MicroScale Thermophoresis.}, author={Dennis Breitsprecher and Nina Schlinck and David Witte and Stefan Duhr and Philipp Baaske and Thomas Schubert}, journal={Methods in molecular biology}, year={2016}, volume.

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