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    A plasmonic gold nanofilm-based microfluidic chip for rapid and inexpensive droplet-based photonic PCR

    , Article Scientific Reports ; Volume 11, Issue 1 , December , 2021 ; 20452322 (ISSN) Jalili, A ; Bagheri, M ; Shamloo, A ; Kazemipour Ashkezari, A. H ; Sharif University of Technology
    Nature Research  2021
    Abstract
    Polymerase chain reaction (PCR) is a powerful tool for nucleic acid amplification and quantification. However, long thermocycling time is a major limitation of the commercial PCR devices in the point-of-care (POC). Herein, we have developed a rapid droplet-based photonic PCR (dpPCR) system, including a gold (Au) nanofilm-based microfluidic chip and a plasmonic photothermal cycler. The chip is fabricated by adding mineral oil to uncured polydimethylsiloxane (PDMS) to suppress droplet evaporation in PDMS microfluidic chips during PCR thermocycling. A PDMS to gold bonding technique using a double-sided adhesive tape is applied to enhance the bonding strength between the oil-added PDMS and the... 

    A triple fouling layers perspective on evaluation of membrane fouling under different scenarios of membrane bioreactor operation

    , Article Journal of Environmental Health Science and Engineering ; Volume 12, Issue 1 , June 2014 Pourabdollah, M ; Torkian, A ; Hashemian, S. J ; Bakhshi, B ; Sharif University of Technology
    Abstract
    One of the main factors affecting membrane fouling in MBRs is operational conditions. In this study the influence of aeration rate, filtration mode, and SRT on hollow fiber membrane fouling was investigated using a triple fouling layers perspective. The sludge microbial population distribution was also determined by PCR method. Through various applied operational scenarios the optimal conditions were: aeration rate of 15 LPM; relaxation mode with 40s duration and 8 min. interval; and SRT of 30 days. The similarity between SMP variations in triple fouling layers with its corresponding hydraulic resistance confirmed the effect of SMP on membrane fouling. Among three fouling fractions, the... 

    Numerical simulation of mixing and heat transfer in an integrated centrifugal microfluidic system for nested-PCR amplification and gene detection

    , Article Sensors and Actuators, B: Chemical ; Volume 283 , 2019 , Pages 831-841 ; 09254005 (ISSN) Naghdloo, A ; Ghazimirsaeed, E ; Shamloo, A ; Sharif University of Technology
    Elsevier B.V  2019
    Abstract
    Nucleic acid amplification via polymerase chain reaction (PCR) is one of the essential and powerful methods used in a myriad of bio-assays in clinical laboratories. Application of microfluidic devices in biologically-related processes like PCR can result in the usage of less volume of reactant samples and reduce the processing time. By implementing PCR systems on centrifugal microfluidic platforms, automation and portability can be easily achieved. Although several methods have been developed, most of them are still dealing with challenges of the required high processing time. This study presents the numerical simulation of a fully automated PCR system with the goal of enhancing the mixing... 

    Design, Simulation and Construction of a Rapid Gene Amplification Microfluidic Device Using Polymerase Chain Reaction (PCR) Method

    , M.Sc. Thesis Sharif University of Technology Amadeh, Ali (Author) ; Shamloo, Amir (Supervisor)
    Abstract
    Polymerase chain reaction method is a conventional method for obtaining multiple copies of a specified segment in the DNA molecule and to amplify the DNA molecule itself. Therefore, this method has been of paramount importance in different fields of research and has been applied for different applications. PCR requires thermal cycling, or repeated temperature changes between two or three discrete temperatures to amplify specific nucleic acid target sequences. To achieve such thermal cycling, conventional bench-top thermal cyclers generally use a metal heating block powered by Peltier elements or benefit from forced convection heat transfer. Although efforts have been made in order to reduce... 

    Design and Fabrication of a Microfluidic Digital PCR

    , M.Sc. Thesis Sharif University of Technology Bahrami, Mohammad Amin (Author) ; Saeedi, Mohammad Saeed (Supervisor) ; Firouzabadi, Bahar (Supervisor)
    Abstract
    Polymerase chain reaction, abbreviated as PCR, is a method of amplifying the number of copies of a desired DNA sequence through special protocols. The rapid advance of microfluidic devices and the emerging concept of digital microfluidics has resulted into the development of digital droplet PCR (ddPCR) systems with their significant uses in the detection of rare mutations, cancer diagnosis, and surveillance. In this study, an integrated ddPCR microfluidic chip alongside its thermoelectric thermal cycler and optical analysis systems have been designed, fabricated and their performance has been tested and evaluated. It was shown that the effect of master mold’s surface quality, the material of... 

    Design of a Microfluidic Digital Droplet PCR

    , M.Sc. Thesis Sharif University of Technology Abedini, Ali (Author) ; Saeedi, Mohammad Saeed (Supervisor)
    Abstract
    Polymerase chain reaction, abbreviated as PCR, is a method of amplifying the number of copies of a desired DNA sequence through special protocols. The rapid advance of microfluidic devices and the emerging concept of digital microfluidics has resulted into the development of digital droplet PCR (ddPCR) systems with their significant uses in the detection of rare mutations, cancer diagnosis, and surveillance. A large number of micron-sized droplets are required to perform ddPCR. in this study, To investigate the gradient of confinements induced droplet self-breakup mechanism, we carried out the computational fluid dynamics (CFD) simulations for the two-phase flow using a commercial software... 

    Cloning and Production of the Flavin Reductase Enzyme in Order to Perform Organic Reaction and Synthesis of Cation Exchange Column for Protein Purification

    , M.Sc. Thesis Sharif University of Technology Hayati, Fatemeh (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    The reductase enzymes perform an important function in the cell. These enzymes are usually coupled with other enzymes and participate in reactions such as halogenation, reduction of alkene, and carboxylic acids. These reactions are carried out by reproducing cofactors such as FADH2 and NAD+; however, there is no evidence that this enzyme alone can perform an organic reaction. In this work, we aim to examine whether recombinant flavin reductase can carry out organic reactions. In order to amplify the flavin reductase enzyme gene from the E. coli genome by PCR (Polymerase Chain Reaction) method, two synthetic primers were designed; The forward primer of the gene contained a cleavage site for... 

    Study, Optimization and Construction of a Microfluidic Gene Amplification Device by Using Thin Film Layer Method

    , M.Sc. Thesis Sharif University of Technology Eslami, Sara Sadat (Author) ; Vosoughi, Manouchehr (Supervisor) ; Alemzadeh, Iran (Co-Supervisor) ; Shamloo, Amir (Co-Supervisor)
    Abstract
    Polymerase Chain Reaction (PCR) is a process in which a special piece of a gene is amplified millions of times over a short period. This method has been of paramount importance in different fields of research and has been applied for different applications. PCR requires thermal cycling, or repeated temperature changes between two or three discrete temperatures to amplify specific nucleic acid target sequences. To achieve such thermal cycling, conventional bench-top thermal cyclers generally use a metal heating block powered by Peltier elements or benefit from forced convection heat transfer. Due to the fact that these methods are time consuming, it seems that design and fabrication of a fast... 

    PCR Amplification Prediction using Machine Learning

    , M.Sc. Thesis Sharif University of Technology Latifian, Niloofar (Author) ; Hossein Khalaj, Babak (Supervisor)
    Abstract
    Polymerase Chain Reaction (PCR) is a laboratory method for amplifying a part of DNA. This method is used in determining the sequence of genes, detecting pathogenic agents in epidemics, creating genetic changes in bacteria, diseases, plants and even animals. Many factors affect the quality of the reaction. Each of these factors can be effective in amplifying the target in DNA. If we can predict the result of PCR using the factors involved in the reaction, it will save a lot of money and time. The aim of this research is to predict the result of PCR amplification using machine learning methods. For this purpose, two methods are proposed: feature-based method and string-based method. In the... 

    Embryonic stem cells maintain an undifferentiated state on dendrimer-immobilized surface with D-glucose display

    , Article Polymers ; Volume 3, Issue 4 , 2011 , Pages 2078-2087 ; 20734360 (ISSN) Mashayekhan, S ; Kim, M. H ; Kino Oka, M ; Miyazaki, J. I ; Taya, M ; Sharif University of Technology
    2011
    Abstract
    In serial passaging cultures of mouse embryonic stem (ES) cells, we employed a dendrimer-immobilized substrate that displayed D-glucose as a terminal ligand. The D-glucose-displaying dendrimer (GLU/D) surface caused the ES cells to form loosely attached spherical colonies, while those on a gelatin-coated surface formed flatter colonies that were firmly attached to the surface. Despite the morphological similarities between the colonies on the GLU/D surface and aggregates on a conventional bacteriological dish, immunostaining and RT-PCR analyses revealed the maintenance of cells within the spherical colonies on the GLU/D surface in an undifferentiated state with very low expressions of... 

    Creating Random Mutations on The Human Transglutaminase 2 Gene and Lysozyme Gene to Increase The Solubility of the Enzymes in order to Perform Promiscuous Organic Reactions Using The Recombinant Enzymes

    , M.Sc. Thesis Sharif University of Technology Moghaddasi, Ahmad (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    Transglutaminase 2 enzyme is expressed in various parts of the human body and is present in almost all tissue cells. The function of this enzyme is multiple and it catalyzes different reactions depending on where it is located in the cell. The most important activities of this enzyme are: isopeptide bond formation between the side chains of two proteins (transamidation reaction), diamidation reaction (hydrolysis of amide bond), esterification, kinase activity. The hen egg white lysozyme protein is another natural antimicrobial protein that has been extensively studied. The muramidase activity of egg white lysozyme against highly gram-positive bacteria is well known, and this property has... 

    Design and fabrication of a two-phase diamond nanoparticle aided fast PCR device

    , Article Analytica Chimica Acta ; Volume 1068 , 2019 , Pages 28-40 ; 00032670 (ISSN) Madadelahi, M ; Ghazimirsaeed, E ; Shamloo, A ; Sharif University of Technology
    Elsevier B.V  2019
    Abstract
    Polymerase Chain Reaction (PCR) is an important and prevalent technique in biotechnology because of its crucial role in cloning DNA fragments and diagnostic applications. In the present study, a high-throughput two-phase PCR device is designed and fabricated which utilizes a serpentine microchannel together with a spiral structure. The former is for the droplet-generation and mixing and the latter is for the thermal cycling process. Moreover, the effect of diamond nanoparticles (diamondNP) on the performance of PCR is also investigated while using commercial PCR devices and the fabricated PCR device designed in this study. Using numerical simulation, it is shown that within the simple and... 

    Improving the performance of a photonic PCR system using TiO2 nanoparticles

    , Article Journal of Industrial and Engineering Chemistry ; 2020 Amadeh, A ; Ghazimirsaeed, E ; Shamloo, A ; Dizani, M ; Sharif University of Technology
    Korean Society of Industrial Engineering Chemistry  2020
    Abstract
    Nucleic acid amplification using polymerase chain reaction (PCR) method has been widely used in different fields such as agricultural science, medicine, pathogen identification, and forensics to name a few. Today, it seems inevitable to have a robust, simple PCR system for diagnostics at the point-of-care (POC) level. Many photonic PCR systems have been proposed in the literature that benefit from plasmonic photothermal heating to achieve the common PCR thermal cycling. However, non-homogeneous temperature distribution is a challenge in some of them. In the present work, to achieve more efficient gene amplification, the effect of adding TiO2 nanoparticles has been investigated in a photonic... 

    Biodesulfurization of dibenzothiophene by a newly isolated Rhodococcus erythropolis strain

    , Article Bioresource Technology ; Volume 101, Issue 3 , 2010 , Pages 1102-1105 ; 09608524 (ISSN) Davoodi Dehaghani, F ; Vosoughi, M ; Ziaee, A. A ; Sharif University of Technology
    2010
    Abstract
    A new dibenzothiophene (DBT) desulfurizing bacterium was isolated from oil-contaminated soils in Iran. HPLC analysis and PCR-based detection of the presence of the DBT desulfurization genes (dszA, dszB and dszC) indicate that this strain converts DBT to 2-hydroxybiphenyl (2-HBP) via the 4S pathway. The strain, identified as Rhodococcus erythropolis SHT87, can utilize DBT, dibenzothiophene sulfone, thiophene, 2-methylthiophene and dimethylsulfoxide as a sole sulfur source for growth at 30 °C. The maximum specific desulfurization activity of strain SHT87 resting cells in aqueous and biphasic organic-aqueous systems at 30 °C was determined to be 0.36 and 0.47 μmol 2-HBP min-1 (g dry cell)-1,... 

    Biological removal of nutrients (N & P) from urban wastewater with a modified integrated fixed-film activated sludge-oxic settling anoxic system using an anoxic sludge holding tank

    , Article Water and Environment Journal ; Volume 35, Issue 2 , 2021 , Pages 830-846 ; 17476585 (ISSN) Fazelipour, M ; Takdastan, A ; Borghei, S. M ; Sharif University of Technology
    John Wiley and Sons Inc  2021
    Abstract
    In this research, the efficiency of the integrated fixed-film activated sludge-oxic settling anoxic (IFAS-OSA) system in biological nutrient removal was studied. The oxic-settling anoxic (OSA) process is known as a cost effective way to reduce the nutrients (nitrogen and Phosphorus). According to the results, the percentages of total nitrogen removal efficiency in the IFAS, IFAS-OSA2h and IFAS-OSA4h systems were 78.56 ± 2.46, 83.60 ± 0.92 and 85.03 ± 1.69, respectively, while the percentages of phosphorus removal efficiency in these systems were 32.69 ± 8.25, 36.35 ± 6.73 and 39.87 ± 3.61, respectively. The PCR-RFLP method showed that C. albicans had the greatest prevalence (n = 36, 90%)....