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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... 

    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... 

    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... 

    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...