Tuesday, June 26, 2018

Nature Methods Application Notes e-UPDATE: 26 June 2018

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26 June 2018 
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FEATURED APPLICATION NOTE
Miniaturized 16S rRNA Amplicon Sequencing with the Labcyte Echo® 525 Liquid Handler for Metagenomic and Microbiome Studies
www.labcyte.com >
Our understanding of the microbial communities in human health, metagenomics, and metataxonomics has been growing rapidly in recent years and is being increasingly elucidated every day. Amplicon sequencing of highly conserved prokaryotic 16S ribosomal RNA (rRNA) regions has long been the standard technique used to assess the diversity and phylogenetic classification of these communities. While advances in next-generation sequencing are enabling routine whole-genome shotgun sequencing in microbial communities, 16S rRNA amplicon sequencing is still frequently used for quick diagnosis of samples. Here, we perform a standard Illumina 16S rRNA amplicon sequencing library preparation at miniaturized scale using the Labcyte Echo® 525 Liquid Handler, effectively reducing reaction volumes and input sample while maintaining sufficient read depth to accurately capture the community. The result is a significantly cost-reduced workflow per sample, saving reagent costs, operational time costs, and valuable sample DNA.
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TwistAmp® Liquid: a versatile amplification method to replace PCR
www.twistdx.co.uk/en/rpa >
Here we introduce TwistAmp® Liquid, a new PCR replacement format that makes RPA technology more amenable to a wide range of research applications. In contrast to PCR amplification, RPA takes minutes, rather than hours, and can be run with little to no equipment. TwistAmp® Liquid Basic and Basic RT kits can be used for applications requiring good fidelity, gel electrophoresis or solid phase detection. TwistAmp® Liquid exo and exo RT allow rapid real-time amplification and detection of targets without sacrificing sensitivity or specificity.
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Acoustic Force Spectroscopy measurements of protein (un)folding at the single-protein level
https://lumicks.com/ >
Single-molecule force spectroscopy (SMFS) technologies have become prominent as powerful tools for the investigation of the biomechanics associated with protein structure and for the study of protein unfolding pathways. In turn, the information accessed by SMFS tools provides valuable contributions to research and drug development. In particular, SMFS can aid in revealing the molecular mechanisms underlying a wide range of human pathologies that are believed to stem from the formation and aggregation of misfolded proteins, such as Alzheimer's and Parkinson's disease.
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Controlling the topography and biochemistry of cell culture substrates with PRIMO® photopatterning system
www.alveolelab.com >
In vivo, the cellular microenvironment has a crucial impact on the regulation of cell behavior and functions such as cellular differentiation, proliferation and migration. One of the challenges confronting cell biologists is to mimic this microenvironment in vivo to more efficiently study living cells and model diseases. Here we present PRIMO: a contactless and maskless UV projection system, which allows to control the topography and chemistry of in vivo microenvironments. We first show that PRIMO is a suitable tool to structure photosensitive resists and create molds on which elastomeric solutions (PDMS) can be polymerized. Then we show that the structured PDMS can be specifically functionalized with biomolecules using UV-light structured by PRIMO and a specific photo-initiator (PLPP™). Altogether, we demonstrate that PRIMO allows to tailor the cell microenvironment topography through microfabrication and biochemistry through micropatterning.
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Anti-idiotypic binders for Trastuzumab validated in regulatory bioanalysis assay in collaboration with Covance
www.avacta.com/ >
Regulatory bioanalysis of therapeutic proteins during drug development and clinical follow-up requires the use of critical reagents that can specifically identify and accurately quantify each biotherapeutic within patient samples. Antibodies currently represent the 'gold standard' of affinity reagents used in the regulated bioanalysis of therapeutic proteins. While traditional antibodies have been refined to the point where they are specific, sensitive and reasonably reliable, they can be limited by their development speed, complexity to produce on an industrial scale and lot-to-lot variation in assay performance.
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Triplex Crystal Digital™ PCR assay to detect HER2 Copy Number amplification.
https://www.stillatechnologies.com >
Investigating HER2 amplification status is crucial in breast cancer to determine the indication of anti-HER2 targeted therapy. Assessing HER2 copy number variation is challenging especially when tumor DNA is diluted in DNA from healthy cells. Here, we illustrate how Crystal Digital PCR is capable of reliably identifying HER2 amplification in samples with low tumoral fraction.
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Rapid and real-time analysis of polyelectrolyte multilayer films using SEEC
http://www.nano-lane.com/ >
Polyelectrolyte multilayers offer novel biosynthetic interfaces that aid in high impact scientific advances in fields such as bioengineering. Current biosensor techniques employed to study the engineered biosynthetic surfaces do not offer real-time visualisation of the layer build-up process and/or surface characterisation studies such as topographic/morphologic analyses. SEEC technology offers enhanced surface contrast that permits quantification of nanometric changes on surfaces such as thin films in liquid conditions. We show that the newly automated N-Lab Station based on SEEC technology offers robust data reproducibility and comprehensive studies including surface characterisation of polyelectrolyte multilayers.
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