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Analytical Sciences : the International... Jul 2021TRIzol is a monophasic solution of phenol and guanidine isothiocyanate used for the extraction of RNA, DNA and proteins from tissues or cells. However, few studies have...
TRIzol is a monophasic solution of phenol and guanidine isothiocyanate used for the extraction of RNA, DNA and proteins from tissues or cells. However, few studies have described its application to DNA extraction due to its time-consuming procedure. We present a TRIzol-modified method of extracting DNA from tissues using the TRIzol reagent and a silica column, which requires only one-third of the time required for the classic extraction procedure. Spectrophotometric analysis showed that the 260/280 and 260/230 nm optical density ratio of the DNA extracted using the TRIzol-modified method is ideal and equal to that obtained by the classic method and commercial DNAiso methods. The DNA extracted by the TRIzol-modified method had the same performance in a restriction enzyme digestion and quantitative PCR as that extracted using the classic method. Using the TRIzol-modified method saves time, simplifies the DNA extraction procedure, and facilitates various molecular biology assays.
Topics: DNA; Guanidines; Indicators and Reagents; Phenols; Silicon Dioxide
PubMed: 33250452
DOI: 10.2116/analsci.20P361 -
BMC Biology Nov 2014The study of microbial communities has been revolutionised in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene...
BACKGROUND
The study of microbial communities has been revolutionised in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. One potential confounder of these sequence-based approaches is the presence of contamination in DNA extraction kits and other laboratory reagents.
RESULTS
In this study we demonstrate that contaminating DNA is ubiquitous in commonly used DNA extraction kits and other laboratory reagents, varies greatly in composition between different kits and kit batches, and that this contamination critically impacts results obtained from samples containing a low microbial biomass. Contamination impacts both PCR-based 16S rRNA gene surveys and shotgun metagenomics. We provide an extensive list of potential contaminating genera, and guidelines on how to mitigate the effects of contamination.
CONCLUSIONS
These results suggest that caution should be advised when applying sequence-based techniques to the study of microbiota present in low biomass environments. Concurrent sequencing of negative control samples is strongly advised.
Topics: DNA Contamination; Indicators and Reagents; Laboratories; Metagenomics; Microbiota; Polymerase Chain Reaction; RNA, Ribosomal, 16S; Salmonella; Sequence Analysis, DNA
PubMed: 25387460
DOI: 10.1186/s12915-014-0087-z -
Report on Carcinogens : Carcinogen... 2011
Topics: Animals; Carcinogens; Humans; Indicators and Reagents; Laxatives; Lymphoma; Phenolphthalein
PubMed: 21863080
DOI: No ID Found -
Lab on a Chip Aug 2009The SlipChip is a microfluidic device designed to perform multiplexed microfluidic reactions without pumps or valves. The device has two plates in close contact. The...
The SlipChip is a microfluidic device designed to perform multiplexed microfluidic reactions without pumps or valves. The device has two plates in close contact. The bottom plate contains wells preloaded with many reagents; in this paper plates with 48 reagents were used. These wells are covered by the top plate that acts as a lid for the wells with reagents. The device also has a fluidic path, composed of ducts in the bottom plate and wells in the top plate, which is connected only when the top and bottom plate are aligned in a specific configuration. Sample can be added into the fluidic path, filling both wells and ducts. Then, the top plate is "slipped", or moved, relative to the bottom plate so the complementary patterns of wells in both plates overlap, exposing the sample-containing wells of the top plate to the reagent-containing wells of the bottom plate, and enabling diffusion and reactions. Between the two plates, a lubricating layer of fluorocarbon was used to facilitate relative motion of the plates. This paper implements this approach on a nanoliter scale using devices fabricated in glass. Stability of preloaded solutions, control of loading, and lack of cross-contamination were tested using fluorescent dyes. Functionality of the device was illustrated via crystallization of a model membrane protein. Fabrication of this device is simple and does not require a bonding step. This device requires no pumps or valves and is applicable to resource-poor settings. Overall, this device should be valuable for multiplexed applications that require exposing one sample to many reagents in small volumes. One may think of the SlipChip as an easy-to-use analogue of a preloaded multi-well plate, or a preloaded liquid-phase microarray.
Topics: Bacterial Proteins; Food Coloring Agents; Hyphomicrobiaceae; Indicators and Reagents; Microfluidic Analytical Techniques; Photosynthetic Reaction Center Complex Proteins
PubMed: 19636458
DOI: 10.1039/b908978k -
Contrast Media & Molecular Imaging 2022As the function and R&D level of in vitro diagnostic reagents continue to improve, the need for hospitals for in vitro diagnostic reagents in clinical diagnosis also...
As the function and R&D level of in vitro diagnostic reagents continue to improve, the need for hospitals for in vitro diagnostic reagents in clinical diagnosis also keeps increasing. However, under the influence of management, process, technology, equipment, materials, employees, and other unexpected disturbing factors, the output of reagents often has random uncertainty, and it is difficult to provide the finished products required by orders on time, in quality and quantity. A secondary supply chain consisting of reagent manufacturers, distributors, and hospitals is constructed, and the inventory control models of in vitro diagnostic reagent supply chain under three strategies of centralized decision-making, hospital-owned inventory, and reagent distributor-managed inventory are established, respectively, and the maximum expected returns of the supply chain system under different strategies are analyzed to achieve the optimal production decision of reagent manufacturers and the optimal procurement decision of hospitals. The results show that reducing the random output probability and patient demand uncertainty has a significant effect on improving the expected return of in vitro diagnostic reagent supply chain, and as the random output probability of reagent manufacturers and patient consumption demand uncertainty increase, the strategy of managing inventory by distributors in collaboration is always better than the strategy of managing inventory by hospitals' own warehouses, which can achieve higher expected return and better inventory quantity, but when the out-of-stock cost of hospitals is too high above a certain threshold, the hospital will tend to adopt the self-inventory strategy.
Topics: Humans; Indicators and Reagents; Pharmaceutical Preparations
PubMed: 35542757
DOI: 10.1155/2022/5046141 -
PloS One 2021We recently developed 'cellular' reagents-lyophilized bacteria overexpressing proteins of interest-that can replace commercial pure enzymes in typical diagnostic and...
We recently developed 'cellular' reagents-lyophilized bacteria overexpressing proteins of interest-that can replace commercial pure enzymes in typical diagnostic and molecular biology reactions. To make cellular reagent technology widely accessible and amenable to local production with minimal instrumentation, we now report a significantly simplified method for preparing cellular reagents that requires only a common bacterial incubator to grow and subsequently dry enzyme-expressing bacteria at 37°C with the aid of inexpensive chemical desiccants. We demonstrate application of such dried cellular reagents in common molecular and synthetic biology processes, such as PCR, qPCR, reverse transcription, isothermal amplification, and Golden Gate DNA assembly, in building easy-to-use testing kits, and in rapid reagent production for meeting extraordinary diagnostic demands such as those being faced in the ongoing SARS-CoV-2 pandemic. Furthermore, we demonstrate feasibility of local production by successfully implementing this minimized procedure and preparing cellular reagents in several countries, including the United Kingdom, Cameroon, and Ghana. Our results demonstrate possibilities for readily scalable local and distributed reagent production, and further instantiate the opportunities available via synthetic biology in general.
Topics: COVID-19; COVID-19 Testing; Cameroon; Diagnostic Tests, Routine; Escherichia coli; Gene Expression; Geobacillus stearothermophilus; Ghana; Humans; Indicators and Reagents; Molecular Diagnostic Techniques; Plasmids; Real-Time Polymerase Chain Reaction; Recombinant Proteins; SARS-CoV-2; Synthetic Biology; Transformation, Bacterial; United Kingdom
PubMed: 34061896
DOI: 10.1371/journal.pone.0252507 -
Bioanalysis Apr 2022To mitigate assay interference in the drug and target assays to support the development of monoclonal antibody REGN-Z. Mild acidic assay conditions and capture and...
To mitigate assay interference in the drug and target assays to support the development of monoclonal antibody REGN-Z. Mild acidic assay conditions and capture and detection antibodies with different affinities and t under different assay pHs were used to mitigate interference in the total drug and total target assays. A free target assay was also developed using a lower-affinity capture antibody with a much slower association and dissociation rate. The impact of sample incubation, dilution and storage on the accurate detection of the free target was also evaluated. The total drug, total and free target assays can accurately quantitate drug and target concentrations when tested with a subset of clinical study samples.
Topics: Antibodies, Monoclonal; Biological Assay; Indicators and Reagents
PubMed: 35297286
DOI: 10.4155/bio-2021-0276 -
IARC Monographs on the Evaluation of... 2000
Review
Topics: Animals; Carcinogenicity Tests; Carcinogens; Cricetinae; Disease Models, Animal; Environmental Exposure; Female; Humans; Indicators and Reagents; Male; Maximum Allowable Concentration; Mice; Neoplasms; Rats; Toluidines
PubMed: 11100404
DOI: No ID Found -
Molecules (Basel, Switzerland) Dec 2022Synthesis of '-Di-Boc-2H-isoindole-2-carboxamidine, the first representative of isoindoles containing guanidine functionality, was carried out. The cycloaddition...
Synthesis of '-Di-Boc-2H-isoindole-2-carboxamidine, the first representative of isoindoles containing guanidine functionality, was carried out. The cycloaddition reactivity of this new Diels-Alder heterodiene was studied and the title compound was employed as a cycloaddition delivery reagent for guanidine functionality. Higher reactivity was found in comparison with the corresponding pyrrole derivative. Substitution with fluorine or guanidine functionality does not change the reactivities of isoindoles, and these findings are in good accord with computational results.
Topics: Guanidine; Isoindoles; Guanidines; Indicators and Reagents; Cycloaddition Reaction
PubMed: 36558087
DOI: 10.3390/molecules27248954 -
ACS Chemical Biology Dec 2016Diazo groups have broad and tunable reactivity. That and other attributes endow diazo compounds with the potential to be valuable reagents for chemical biologists. The... (Review)
Review
Diazo groups have broad and tunable reactivity. That and other attributes endow diazo compounds with the potential to be valuable reagents for chemical biologists. The presence of diazo groups in natural products underscores their metabolic stability and anticipates their utility in a biological context. The chemoselectivity of diazo groups, even in the presence of azido groups, presents many opportunities. Already, diazo compounds have served as chemical probes and elicited novel modifications of proteins and nucleic acids. Here, we review advances that have facilitated the chemical synthesis of diazo compounds, and we highlight applications of diazo compounds in the detection and modification of biomolecules.
Topics: Alkylation; Amino Acids; Animals; Biochemistry; Biological Products; Cycloaddition Reaction; Diazonium Compounds; Humans; Indicators and Reagents; Models, Molecular; Nucleic Acids; Proteins
PubMed: 27739661
DOI: 10.1021/acschembio.6b00810