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Real-time isothermal DNA amplification monitoring in picoliter volumes using an optical fiber sensor
PublikacjaRolling circle amplification (RCA) of DNA can be considered as a great alternative to the gold standard polymerase chain reaction (PCR), especially during this pandemic period, where rapid, sensitive, and reliable test results for hundreds of thousands of samples are required daily. This work presents the first research to date on direct, real-time and label-free isothermal DNA amplification monitoring using a microcavity in-line...
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Discovery and characterization of RecA protein of thermophilic bacterium Thermus thermophilus MAT72 phage Tt72 that increases specificity of a PCR-based DNA amplification
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From Individual to Collective: Intelligence Amplification with Bio-Inspired Decisional DNA and its Extensions
PublikacjaIn nature, deoxyribonucleic acid (DNA) contains the genetic instructions used in the development and functioning of all known living organisms. The idea behind our vision is to develop an artificial system, an architecture that would support discovering, adding, storing, improving and sharing information and knowledge among agents and organizations through experience. We propose a novel Knowledge Representation (KR) approach in...
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A vector-enzymatic DNA fragment amplification-expression technology for construction of artificial, concatemeric DNA, RNA and proteins for novel biomaterials, biomedical and industrial applications
PublikacjaA DNA fragment amplification/expression technology for the production of new generation biomaterials for scientific, industrial and biomedical applications is described. The technology enables the formation of artificial Open Reading Frames (ORFs) encoding concatemeric RNAs and proteins. It recruits the Type IIS SapI restriction endonuclease (REase) for an assembling of DNA fragments in an ordered head-to-tail-orientation. The...
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Evaluation and comparison of random amplification of polymorphic DNA, pulsed field gel electrophoresis and ADSRRS-fingerprinting for typing Serratiamarcescens outbreaks.
PublikacjaADSRRS-fingerprinting, nowa metoda genotypowania oparta na supresji reakcji PCR została zastosowana do badań epidemiologicznych izolatów S. marcescens,pochodzących z trzech różnych epidemii Szpitala Publicznego w Gdańsku. Wyniki uzyskane metodą ADSRRS-fingerprinting porównano z wynikami uzyskanymi technikami RAPD-fingerprinting i PFGE.
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Data regarding a new, vector-enzymatic DNA fragment amplification-expression technology for the construction of artificial, concatemeric DNA, RNA and proteins, as well as biological effects of selected polypeptides obtained using this method
PublikacjaApplications of bioactive peptides and polypeptides are emerging in areas such as drug development and drug delivery systems. These compounds are bioactive, biocompatible and represent a wide range of chemical properties, enabling further adjustments of obtained biomaterials. However, delivering large quantities of peptide derivatives is still challenging. Several methods have been developed for the production of concatemers –...
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Evaluation of a novel method based on amplification of DNA fragments surrounding rare restriction sites (ADSRRS-figerprinting) for typing strains of vancomycin resistant Enterococcus faecium.
PublikacjaBadania objęły wankomycyno oporne szczepy Enterococcus faecium izolowane od pacjentów oddziału Hematologii Szpitala Klinicznego w Gdańsku. Wykazano, że ADSRRS-fingerprinting jest metodą szybką o dużym potencjale różnicującym i dużej powtarzalności w badaniach epidemiologicznych.
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Loop-mediated isothermal amplification (LAMP) as a diagnostic tool in detection of infectious diseases
PublikacjaLoop-mediated isothermal amplification (LAMP) is a gene amplification method which amplifies DNA with high specificity and efficiency under isothermal conditions. Because of its rapidity and simplicity, it is a valuable diagnostic tool in the early detection and identification of infectious diseases. LAMP method is based on the use of a set of four to six specially designed primers spanning six to eight distinct sequences on the...
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Principles and applications of Ligation Mediated PCR methods for DNA-based typing of microbial organisms
PublikacjaA significant number of DNA-based techniques has been introduced into the field of microorganisms’ characterization and taxonomy. These genomic fingerprinting methods were developed to detect DNA sequence polymorphisms by using general principles, such as restriction endonuclease analysis, molecular hybridization, and PCR amplification. In recent years, some alternative techniques based on ligation of oligonucleotide adapters before...
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Taq DNA polymerase fused with DNA binding protein with increased resistance to inhibitors from clinical samples
PublikacjaNowadays PCR method is commonly used in molecular diagnostic. However, in many cases PCR is limited, by the presence of inhibitory substances in biological, soil or food samples Efficiency and fidelity of amplification is strongly connected with DNA polymerase and reaction conditions. To meet the requirements of modern diagnostic methods it is essential to seeking for new DNA polymerases with better properties useful in these field....
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Increased concentration of Taq DNA polymerase as a solution for GC-rich templates from clinical and environmental samples
PublikacjaDNA polymerase is an enzyme which plays crucial role in replication and DNA repair. It found application in PCR (polymerase chain reaction) where catalyses process of in vitro DNA synthesis. To meet the demands posed by mod- ern diagnostic, molecular biology or genetic engineering it is necessary to improve DNA polymerases to obtain new or better features useful in these fields. So far implemented modifications in majority are...
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Fuzyjne polimerazy DNA – otrzymywanie, charakterystyka i zastosowanie
PublikacjaObecnie reakcje PCR (ang. Polymerase Chain Reaction) wykazują bardzo szerokie zastosowanie w diagnostyce medycznej, biologii molekularnej czy inżynierii genetycznej. Efektywność tych reakcji rozumiana jako wydajność i wierność przeprowadzonej amplifikacji jest nieodłącznie związana ze stosowaną polimerazą DNA i warunkami prowadzenia reakcji PCR. Aby sprostać wymaganiom stawianym przez nowoczesne metody diagnostyczne oraz współczesną...
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Experience-based Intelligence Augmentation with Decisional DNA: Upcoming direction
PublikacjaIntelligence amplification systems and technologies have gained significant interest from academia and industry during the past few decades. One of the main reasons behind this trend is the fact that most experts agree that truly intelligent artificial system is yet to be developed. The question increasing often asked is this: Is full replication of human intelligence desirable key aim in intelligence related technology and research?...
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Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans—Expression and characterization
PublikacjaDNA polymerases are present in all organisms and are important enzymes that synthesise DNA molecules. They are used in various fields of science, predominantly as essential components for in vitro DNA syntheses, known as PCR. Modern diagnostics, molecular biology and genetic engineering need DNA polymerases which demonstrate improved performance. This study was aimed at obtaining a new NeqSSB-TaqS fusion DNA polymerase from the...
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D-loop sequences retrieved from Canis lupus familiaris mitochondrial genome
Dane BadawczeCanine mitochondrial genome is built of 16727 bp. Non-coding control region (mtCR), called also D-loop, begins with 15458 nucleotide and ends with 16727 nucleotide. The length of this fragment is 1270 bp (Kim et al., 1998). D-loop region is responsible for replication and transcription of mitochondrial DNA. Mutations that occur within it may cause irregularity...
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Typing of Candida isolates from patients with invasive infection and concomitant colonization
PublikacjaWe investigated the relationship between colonizing and invasive isolates from patients with candidaemia. Molecular typing was performed using random amplification of polymorphic DNA (RAPD) and multilocus sequence typing (MLST). We found MLST to be sufficient for typing Candida isolates, and that surveillance cultures are helpful in predicting concomitant invasive isolates, but not necessarily the pathogen involved in subsequent...
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Teloxantron inhibits the processivity of telomerase with preferential DNA damage on telomeres
PublikacjaTelomerase reactivation is one of the hallmarks of cancer, which plays an important role in cellular immortalization and the development and progression of the tumor. Chemical telomerase inhibitors have been shown to trigger replicative senescence and apoptotic cell death both in vitro and in vivo. Due to its upregulation in various cancers, telomerase is considered a potential target in cancer therapy. In this study, we identified...
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ALIS-FLP: amplified ligation selected fragment-length polymorphism method for microbial genotyping
PublikacjaA DNA fingerprinting method known as ALIS-FLP (amplified ligation selected fragment-length polymorphism) has been developed for selective and specific amplification of restriction fragments from TspRI restriction endonuclease digested genomic DNA. The method is similar to AFLP, but differs in that only one specific restriction enzyme (TspRI) is used. The cohesive ends of the DNA fragments are ligated with two types of oligonucleotide....
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Recombinant !ermostable AP Exonuclease from Thermoanaerobacter tengcongensis: Cloning, Expression, Purification, Properties and PCR Application
PublikacjaApurinic/apyrimidinic (AP) sites in DNA are considered to be highly mutagenic and must be corrected to preserve genetic integrity, especially at high temperatures. !e gene encoding a homologue of AP exonuclease was cloned from the thermophilic anaerobic bacterium Thermoanaerobacter tengcongensis and transformed into Escherichia coli. The protein product showed high identity (80%) to human Ape1 nuclease, whereas to E. coli exonuclease...
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Real-Time PCR: molecular technique of many applications
PublikacjaReal-Time PCR is a sensitive DNA amplification technique initially applied in genetics and molecular biology. It enables in vivo copying of the selected DNA fragment (flanked by two primers) by the thermostable polymerase (in the presence of magnesium ions and deoxynucleotide triphosphates) and simultaneous measurement of the fluorescence. For one or more specific sequences in a DNA sample, real-time PCR enables both detection...
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The use of a one-step PCR method for the identification of Microsporum canis and Trichophyton mentagrophytes infection of pets
PublikacjaIntroduction: Dermatophytes are a closely related group of keratinophilic fungi. They encompass important etiological agents of superficial fungal infections. These fungi are able to invade keratinized tissues of humans and animals, causing dermatophytosis (ringworm) of hair, nails or skin. The aim: Traditional diagnostics of ringworm is based on morphological identification of cultured fungi and is time-consuming. Materials and methods:...
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The New LM-PCR/Shifter Method for the Genotyping of Microorganism
PublikacjaTechniques relies on the ligation of appropriates adapters (LM-PCR) as AFLP, PCR MP and ADSRRS are successfully used for epidemiological studies for prokaryotic and eukaryotic microorganisms. In this study we propose a new method, called the LM-PCR/Shifter, based on the use of a Class IIS restriction enzyme giving restriction fragments with different 4 base 5' overhangs (Shifter) and the ligation of appropriate oligonucleotide...
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Genotyping Techniques for Determining the Diversity of Microorganisms
PublikacjaTyping of microbial pathogens, or identifying bacteria at the strain level, is particularly important for diagnosis, treatment, and epidemiological surveillance of bacterial infections. This is especially the case for bacteria exhibiting high levels of antibiotic resistance or virulence, and those involved in nosocomial or pandemic infections. Strain typing also has applications in studying bacterial population dynamics. The part...
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ADSRRS-fingerprinting and PCR MP techniques for studies of intraspecies genetic relatedness in Staphylococcus aureus
PublikacjaW pracy pokazano użyteczność dwóch nowych metod do typowania molekularnego Staphylococcus aureus, ADSRRS-fingerprinting (Amplification of DNA fragments Surrounding Rare Restriction Sites) i PCR MP (PCR melting profile). Przebadano 37 szczepów Staphylococcus aureus izolowanych od pacjentów z czyracznością. Jako metodę referencyjną zastosowano REA-PFGE. Okazało się, że izolaty Staphylococcus aureus z nosa od niektórych pacjentów...
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Optymalizacja i walidacja metody ADSRRS-fingerprinting - konstrukcja zestawu diagnostycznego do różnicowania genetycznego drobnoustrojów
PublikacjaSpośród wszystkich metod typowania mikroorganizmów na szczególną uwagę zasługują metody oparte na analizie całego materiału genetycznego zawartego w komórce. Jedną z nich jest stosunkowo nowa technika ADSRRS-fingerprinting (ang. Amplification of DNA Fragments Surrounding Rare Restriction Sites). Na podstawie naszych badań można stwierdzić, iż charakteryzuje się ona wysokim potencjałem różnicującym, wiarygodnością, powtarzalnością...
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A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification
PublikacjaCandida sp. have been responsible for an increasing number of infections, especially in patients with immunodeficiency. Species specific differentiation of the Candida sp. is difficult in routine diagnosis. This identification can have a highly significant association in therapy and prophylaxis. This work has shown a new application of terminal restriction fragment length polymorphism (t-RFLP) method in molecular identification...
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Investigation of Microbes in the Stratosphere – Stardust Project
PublikacjaThe stratospheric microbiota has been investigated many times using the methods of classical microbiology. In this research a few modern methods, including NGS sequencing and multiple displacement amplification of DNA, were in use. The analysis of metagenome helped to determine the content of various species of bacteria in the sample collected in the stratosphere by a hydrogen-filled balloon holding...
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Evaluation of possibilities of genotyping of Candida glabrata clinical isolates with RAPD-PCR method and microsatellite analysis
PublikacjaThe aim of the study was to compare the discriminatory power of RAPD-PCR method using RSD10 primer and microsatellite (GACA)4 analysis for genotyping clinical isolates of C. glabrata. Isolates were received from patients of two Polish hospitals: Children's Memorial Health Institute in Warsaw (n=17) and Medical University of Gdansk (n=37). Species identification was confirmed by two phenotypic methods: growth on chromagar plates...
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Stardust - Investigation of Microbes in the Stratosphere
PublikacjaThe stratospheric microbiome has been investigated several times using the methods of classical microbiology. In this experiment, we have combined them with some novel approaches including whole- metagenome amplification, Maldi TOF mass spectrometry and Sanger DNA sequencing. The results of the experiment may provide the scientists with knowledge about the mechanisms of survivability of microorganisms in stratospheric conditions...
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Detection of Toxoplasma gondii Infection in Small Ruminants: Old Problems, and Current Solutions
PublikacjaToxoplasmosis is a parasitic zoonosis of veterinary importance, with implications for public health. Toxoplasma gondii infection causes abortion or congenital disease in small ruminants. Moreover, the consumption of infected meat, cured meat products, or unpasteurized milk and dairy products can facilitate zoonotic transmission. Serological studies conducted in various European countries have shown the high seroprevalence of specific...
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Tetracycline and ampicillin resistance of Escherichia coli strains of surface water origin: the potential for horizontal transfer of resistance genes = Oporność na tetracyklinę i ampicylinę szczepów Escherichia Coli wyizolowanych z wody powierzchniowej : możliwość horyzontalnego przenoszenia genów oporności
PublikacjaThe aim of this preliminary study was to assess the occurrence and molecular diversity of tetracycline and ampicillin resistance genes carried by Escherichia coli present in surface water. Bacterial strains were isolated from two watercourses (the Reda River and the Oliwski Stream) that influence the quality of coastal waters in the Gdańsk Bay (Northern Poland) by direct discharge. The bacterial drug susceptibility, tested against...