scholarly journals High-throughput, Automated Extraction of DNA and RNA from Clinical Samples using TruTip Technology on Common Liquid Handling Robots

10.3791/50356 ◽  
2013 ◽  
Author(s):  
Rebecca C. Holmberg ◽  
Alissa Gindlesperger ◽  
Tinsley Stokes ◽  
Dane Brady ◽  
Nitu Thakore ◽  
...  
Keyword(s):  
High Throughput ◽  
Clinical Samples ◽  
Automated Extraction ◽  
Liquid Handling ◽  
Dna And Rna

Automation of a fluorescence-based multiplex PCR for the laboratory confirmation of common bacterial pathogens

2004 ◽  
Vol 53 (2) ◽  
pp. 115-117 ◽  
Author(s):  
Karen Smith ◽  
Mathew A. Diggle ◽  
Stuart C. Clarke
Keyword(s):  
Public Health ◽  
Streptococcus Pneumoniae ◽  
Multiplex Pcr ◽  
High Throughput ◽  
Health Management ◽  
Simultaneous Detection ◽  
Clinical Samples ◽  
Liquid Handling ◽  
Laboratory Confirmation ◽  
Extraction Liquid

A fluorescence-based multiplex PCR was automated for the simultaneous detection of Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae in clinical samples from patients with suspected meningitis. Sensitivity of one to two genome copies per 100 μl sample and specificity of 100 % for each organism were shown. Automation of DNA extraction, liquid handling, PCR and analysis are achieved on a single platform, which enables a high throughput and rapid turnaround of clinical samples that, in turn, leads to faster diagnosis. This is ultimately beneficial to the treatment of the patient and for public health management.

High throughput semi-automated extraction method for the creation of a collection from microbial fermentations of fungi and Actinomycetes

Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
JR Tormo ◽  
N Tabanera ◽  
D Conway ◽  
P Ramos ◽  
A Redondo ◽  
...  
Keyword(s):  
High Throughput ◽  
Extraction Method ◽  
Automated Extraction ◽  
The Creation

scholarly journals A high-throughput screening method for evolving a demethylase enzyme with improved and new functionalities

2020 ◽  
Author(s):  
Yuru Wang ◽  
Christopher D Katanski ◽  
Christopher Watkins ◽  
Jessica N Pan ◽  
Qing Dai ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Method ◽  
Targeted Mutagenesis ◽  
Rna Repair ◽  
Dna And Rna ◽  
Modified Dna ◽  
Dna Substrates ◽  
Trna Sequencing

Abstract AlkB is a DNA/RNA repair enzyme that removes base alkylations such as N1-methyladenosine (m1A) or N3-methylcytosine (m3C) from DNA and RNA. The AlkB enzyme has been used as a critical tool to facilitate tRNA sequencing and identification of mRNA modifications. As a tool, AlkB mutants with better reactivity and new functionalities are highly desired; however, previous identification of such AlkB mutants was based on the classical approach of targeted mutagenesis. Here, we introduce a high-throughput screening method to evaluate libraries of AlkB variants for demethylation activity on RNA and DNA substrates. This method is based on a fluorogenic RNA aptamer with an internal modified RNA/DNA residue which can block reverse transcription or introduce mutations leading to loss of fluorescence inherent in the cDNA product. Demethylation by an AlkB variant eliminates the blockage or mutation thereby restores the fluorescence signals. We applied our screening method to sites D135 and R210 in the Escherichia coli AlkB protein and identified a variant with improved activity beyond a previously known hyperactive mutant toward N1-methylguanosine (m1G) in RNA. We also applied our method to O6-methylguanosine (O6mG) modified DNA substrates and identified candidate AlkB variants with demethylating activity. Our study provides a high-throughput screening method for in vitro evolution of any demethylase enzyme.

scholarly journals A fully automated pipeline for the dynamic at‐line morphology analysis of microscale Aspergillus cultivation

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Roman Jansen ◽  
Kira Küsters ◽  
Holger Morschett ◽  
Wolfgang Wiechert ◽  
Marco Oldiges
Keyword(s):  
Filamentous Fungi ◽  
High Throughput ◽  
Automated Image Analysis ◽  
Height Velocity ◽  
Pellet Size ◽  
Application Study ◽  
Liquid Handling ◽  
Morphology Analysis ◽  
Development And Utilization ◽  
Increasing Demand

Abstract Background Morphology, being one of the key factors influencing productivity of filamentous fungi, is of great interest during bioprocess development. With increasing demand of high-throughput phenotyping technologies for fungi due to the emergence of novel time-efficient genetic engineering technologies, workflows for automated liquid handling combined with high-throughput morphology analysis have to be developed. Results In this study, a protocol allowing for 48 parallel microbioreactor cultivations of Aspergillus carbonarius with non-invasive online signals of backscatter and dissolved oxygen was established. To handle the increased cultivation throughput, the utilized microbioreactor is integrated into a liquid handling platform. During cultivation of filamentous fungi, cell suspensions result in either viscous broths or form pellets with varying size throughout the process. Therefore, tailor-made liquid handling parameters such as aspiration/dispense height, velocity and mixing steps were optimized and validated. Development and utilization of a novel injection station enabled a workflow, where biomass samples are automatically transferred into a flow through chamber fixed under a light microscope. In combination with an automated image analysis concept, this enabled an automated morphology analysis pipeline. The workflow was tested in a first application study, where the projected biomass area was determined at two different cultivation temperatures and compared to the microbioreactor online signals. Conclusions A novel and robust workflow starting from microbioreactor cultivation, automated sample harvest and processing via liquid handling robots up to automated morphology analysis was developed. This protocol enables the determination of projected biomass areas for filamentous fungi in an automated and high-throughput manner. This measurement of morphology can be applied to describe overall pellet size distribution and heterogeneity.

scholarly journals Fast and inexpensive protocols for consistent extraction of high quality DNA and RNA from challenging plant and fungal samples for high-throughput SNP genotyping and sequencing applications

PLoS ONE ◽  
2018 ◽  
Vol 13 (10) ◽  
pp. e0206085 ◽  
Author(s):  
Peter W. Inglis ◽  
Marilia de Castro R. Pappas ◽  
Lucileide V. Resende ◽  
Dario Grattapaglia
Keyword(s):  
High Throughput ◽  
Snp Genotyping ◽  
High Quality ◽  
Dna And Rna

The dispensing well plate: a novel device for nanoliter liquid handling in ultra high-throughput screening

2004 ◽  
Vol 9 (5) ◽  
pp. 291-299 ◽  
Author(s):  
Reinhard Steger ◽  
Benjamin Bohl ◽  
Roland Zengerle ◽  
Peter Koltay
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Novel Device ◽  
Liquid Handling
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