Drug Research
Drug Discovery & Development

Trinean - UV-VIS Spectral Analysis of DNA/RNA, Protein or Small Compounds Case Studies

Trinean - UV-VIS Spectral Analysis of DNA/RNA, Protein or Small Compounds

Sample Processing for the UK Biobank Genotyping Project UK Biobank is a health research charity which has collected and/or generated a wide range of biological samples and data on 500,000 participants aged 40-69 years old (at baseline). Data and samples are available for use by bona-fide researchers to investigate aetiology of disease.
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Precise Nucleic Acid Quantification Using Lab-on-a-chip Spectroscopy and the cDrop Quantification Algorithm Precise quantification and quality control of isolated genomic DNA is critical for the success of downstream applications such as next-generation sequencing. Traditionally, two quantification methods have been used routinely: (1) UV/Vis spectroscopy and (2) fluorescence-based assays (i.e. PicoGreen) that utilize DNA intercalating agents.
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New approach with Spectral Content Profiling allows in-depth plant DNA QC during NGS library preparation As NGS library construction is an expensive and laborious procedure, it is prudent to include quality control steps throughout the library construction process ensuring the quality of downstream data. First, assessing the yield and purity of DNA extracts from plant tissue is challenging due to the fragmented state of the isolated DNA, the presence of co-extracted metabolites (like polyphenolic metabolites, polysaccharides or chlorophylls) and turbidity effects by cell debris or magnetic bead residues. Furthermore, carry-over from the DNA isolation buffers, such as chaotropic salts or phenol can influence the success rate of downstream library preparation or amplification steps.
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Standardizing Protein Quantification of Purified His-tagged Proteins using a Micro-Volume UV/VIS Reader with cDrop Software Purification of recombinant proteins from lysates or supernatants is commonly performed by affinity tagging followed by immobilized metal-affinity chromatography (IMAC). Polyhistidine-tagged (PHT) proteins bind with high affinity to Ni-columns and can subsequently be eluted by adding free imidazole.
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Standardizing the Quality Control of any DNA Isolation to Safeguard the Success of Genetic Testing Genetic tests become ever more powerful, yet also more complex and costly, which is why it is prudent to include quality control steps thoughout the preparation process to ensure the quality of the downstream data obtained.
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All-purpose Protein Sample QC using a Novel Micro-Volume Platform with cDrop Software Accurate total protein quantitation of purified proteins, cell lysates, affinity-column fractions or other complex protein samples is crucial for robust biochemical testing during biopharmaceutical development as well as production. As the presence of chemicals doesn't allow simple A280 protein quantification, more labor-intensive alternatives for quantification are used. Examples are the colorimetric Bradford or BCA assay, which negate to a large extent the presence of contaminants but require multiple steps and show variation depending on the sample composition and time point of measuring within the ongoing reaction.
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DropSense16 Apps - RNA Cy3/Cy5 and ssDNA Cy3/Cy5 An app for specific quantification of Cy3/Cy5 labeled RNA or ssDNA is available on DropSense16. Trinean’s proprietary cDrop software is used to isolate the RNA or ssDNA profile as well as specific Cy3 and Cy5 profiles from the measured UV/VIS spectrum, thereby distinguishing them from other absorbing sample contaminants. These profiles (RNA or ssDNA + Cy3 and Cy5) are then used to determine the total RNA or ssDNA concentration plus amount of Cy3 and Cy5 label. In this note we describe how to use this application and how to interpret its results.
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DropSense16 App – RNA FFPE An app for specific quantification of total RNA extracted from FFPE tissue sections is available on DropSense16. Trinean’s proprietary cDrop software is used to isolate the profile of the desired molecule (here RNA) from the measured UV/VIS spectrum, thereby distinguishing it from absorbing sample contaminants. The specific RNA profile is then used to determine the RNA concentration. In this note we describe how to use this application and how to interpret its results.
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DropSense16 App – Colorimetric assays Often protein quantification is done by measuring absorbance at 280 nm, however results might be influenced by buffer or source constituents. An alternative to A280 quantification are colorimetric protein assays. Reagent(s) and proteins are mixed, producing a colour change which is a measure for the amount of proteins present. Unknown sample concentrations are calculated using a measured standard curve. Apps to measure 4 commonly used colorimetric protein assays are now available on DropSense16.
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DropSense16 App – A280 protein An app for quantification of total purified protein (mg/ml), based on the classic A280 peak height determination. On the DropSense16, apps are available using the classic single-wavelength baseline corrected UV/VIS spectrum (indicated with –bg(340)) or an innovative turbidity full spectral background correction. In this note we describe how to use these applications and how to interpret their results.
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DropSense16 App – RNA A specific app for precise quantification of RNA using Trinean's proprietary cDrop software is available on DropSense16. This advanced analytical method is used to isolate the profile of the desired molecule (here RNA) from the measured UV/VIS spectrum, thereby distinguishing it from interfering chemicals. The specific RNA profile is then used for determining the concentration. In this note we describe how to use this application and how to interpret its results.
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DropSense16 App – Purified PCR An app for specific quantification of DNA amplicon in purified PCR samples is available on the DropSense16. Trinean's proprietary cDrop software is used to isolate the profile of the desired molecule (here DNA) from the measured UV/VIS spectrum, thereby distinguishing it from absorbing sample contaminants. The specific DNA profile is then used to determine the DNA concentration. In this note we describe how to use this application and how to interpret its results.
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DropSense16 App – Culture OD600 In microbiology, optical density measurements at 600 nm (OD600) are used to monitor growth of bacterial cultures. An OD600 application is now available on the DropSense16. The measured UV/VIS spectrum is used to determine the optical density of suspension samples at 600 nm. This note describes how to use this application and how to interpret its results.
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DropSense16 App – DNA mammalian An app for precise quantification of mammalian DNA is available on DropSense16. Trinean's proprietary cDrop software is used to analyse the UV/VIS spectral shape of the sample to isolate the profile of the desired analyte (here DNA) from interfering contaminants. The specific DNA profile is then used to determine the correct DNA concentration. In this note we describe how to use this application and how to interpret its results.
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DropSense16 App – Total Protein An app for total protein quantification using Trinean's proprietary cDrop software is available on the Dropsense16. This advanced analytical method is used to isolate the profile of the desired molecule (here protein) from the measured UV/VIS spectrum, thereby distinguishing it from interfering nucleic acids and chemicals. The specific protein profile is then used for determining its concentration. In this note we describe how to use this application and how to interpret its results.
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