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Potential of Microalgae for Protein Production
Published in Sanjeet Mehariya, Shashi Kant Bhatia, Obulisamy Parthiba Karthikeyan, Algal Biorefineries and the Circular Bioeconomy, 2022
Elena M. Rojo, Alejandro Filipigh, David Moldes, Marisol Vega, Silvia Bolado
The electrophoresis process uses the charge of the molecules to make them migrate through an electric field and is commonly used for protein and other macromolecule separation (Corrêa et al., 2021). The advantage of this approach is that the molecules are first separated based on their charges and then according to their molar masses. This can be achieved by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). It can isolate bands of protein in a simple and relatively rapid manner when used with a staining method (Shen, 2019). Vizcaíno et al. (2019) carried out an electrophoresis with SDS-PAGE to separate different proteins from 4 types of microalgae (Tisochrysis lutea, Nannochloropsis gaditana, Tetraselmis suecica, and Scenedesmus almeriensis) after an enzymatic hydrolysis time of 90min with digestive enzymes from fish extracts at 25°C, obtaining protein bands of 11.7 to 44.5kDa for Tisochrysis lutea, 41 to 57.3kDa for Nannochloropsis gaditana, 24.1 to 55kDa for Tetraselmis suecica, and 26.8 to 149.2 for Scenedesmus almeriensis.
Principles and Techniques for Deoxyribonucleic Acid (DNA) Manipulation
Published in Hajiya Mairo Inuwa, Ifeoma Maureen Ezeonu, Charles Oluwaseun Adetunji, Emmanuel Olufemi Ekundayo, Abubakar Gidado, Abdulrazak B. Ibrahim, Benjamin Ewa Ubi, Medical Biotechnology, Biopharmaceutics, Forensic Science and Bioinformatics, 2022
Nwadiuto (Diuto) Esiobu, Ifeoma M. Ezeonu, Francisca Nwaokorie
DGGE (denatured gradient gel electrophoresis) is a type of gel electrophoresis in which constant heat (about 60°C) and an increasing concentration of denaturing chemicals (in a gel matrix) are used to unwind (i.e. denature) DNA molecules. The single strands are then forced through the varying pores of the vertical electrophoresis system in a process, so powerful it can distinguish single nucleotide polymorphisms (DNA that differs only by one nucleotide) in DNA samples. As already described in preceding sections, electrophoresis techniques separate molecules (DNA, RNA and Protein) on the basis of their electrical charge, shape and molecular weight. In DGGE, DNA is made to migrate through the pores of a polyacrylamide gel of varying concentrations. The DNA unwinds (denatures or melts) as it reaches the denaturing reagents, allowing differential sequences to migrate according to the size and base composition of the fragment. DGGE applications include distinguishing between mutated and wild type sequences without prior knowledge of what these sequences are, detection of mutations within closely related organisms and also organismal differences within an ecosystem. Using SNP detections, DGGE can be used in cloning analysis and cancer cell detection. The resulting gel picture bands can be analyzed using software such as Gel Compar II or Bionumerics to provide statistical analysis and UPGMA clustering profiles.
Capillary Electrophoresis
Published in Grinberg Nelu, Rodriguez Sonia, Ewing’s Analytical Instrumentation Handbook, Fourth Edition, 2019
Electrophoresis is a technique that separates charged molecules based on their movement through a fluid matrix under an applied electric field. The history of electrophoresis has been published by Vesterberg [1] and Compton and Brownlee [2]. The first modern capillary electrophoresis (CE) technique developed was isotachophoresis in 1979 [3] followed by capillary zone electrophoresis (CZE) in the early 1980s [4, 5]. Jorgenson and Lukacs [6] provided breakthroughs in injection and detection using 75 µm i.d. capillaries. Their work was instrumental in defining the start of high-performance capillary electrophoresis (HPCE). Terabe et al. [7] first described micellar electrokinetic capillary chromatography (MEKC), in which chromatographic separations of charged or neutral small molecules were obtained by employing a micelle as a “pseudo-stationary” phase. The first fully automated commercial instrument with an ultraviolet-visible (UV-Vis) detector was introduced in 1989 by Beckman. Many other companies have developed commercial instruments since this time with many fully automated features in HPCE to streamline method development and allow routine use in the laboratory.
Bioprocessing of recombinant proteins from Escherichia coli inclusion bodies: insights from structure-function relationship for novel applications
Published in Preparative Biochemistry & Biotechnology, 2023
Kajal Kachhawaha, Santanu Singh, Khyati Joshi, Priyanka Nain, Sumit K. Singh
Capillary electrophoresis, also called high-efficiency capillary electrophoresis, is a class of liquid phase micro-separation analysis method. It is used for the separation of ions on the basis of their electrophoretic mobility under an applied voltage. The mobility of these ions depends upon the molecule charge, size and viscosity. The mobility of the molecules is directly proportional to the applied electric field. Capillary electrophoresis is utilized very commonly due to faster results and high-resolution separation.[166] Garza et al. developed an IB solubilization method and determined the purity of recombinant proteins in inclusion bodies using capillary gel electrophoresis. It is based on the utilization of a single-component solution which is completely compatible with capillary gel electrophoresis analysis. In this method, IBs sample were prepared and separated from recombinant proteins and other impurities. This allows capillary gel electrophoresis to be regarded as a suitable analytical tool to obtain the quantitative process information during IB refolding.[167]
A shape factor model for injection analysis of microchip sample electrophoresis
Published in Numerical Heat Transfer, Part A: Applications, 2020
Pegah Pezeshkpour, Gerry Schneider, Carolyn Ren
Electrophoresis, which refers to electric field and migration, is applied for separation and characterization of charged particles and macromolecules such as DNA, proteins, RNA, and peptides through a solvent under an applied electric field. The miniaturization of traditional, cumbersome laboratory equipment onto microchip devices, offers the potential for decreased analysis times, reduced sample volumes, reduced operating and manufacturing costs, as well as portability. However, such downsizing necessitates a fundamental study of microscale fluid transportation, microchip design, channel geometries, sample manipulation, and detection methods. A 3 D numerical model for the equilibrium shape of a droplet in a uniform electric field in bubble separation and deformation are presented [1] and the effect of channel geometry on the flow field was also numerically studied in another research [2].
Evolutionary DNA Computing Algorithm for Job Scheduling Problem
Published in IETE Journal of Research, 2018
Gudar J. Ibrahim, Tarik A. Rashid, Ahmed T. Sadiq
For that reason, a Watson–Crick complementary string can be produced via a Turing machine which can code various programs to emulate games such as chess, etc. The basic DNA computing operations that are widely used can be described as follows [23,34,36,38,41–49]: Pairing of Watson–Crick: this prevalent, clearly, strands of DNA can have their Watson–Crick string complements. If a DNA molecule comes across Watson–Crick original complementary strand, the process of pairing will take place. Therefore, both DNA strands can get joined and annealed to yield the double helix.Polymerases: DNA polymerases can generate DNA complementary strands to Watson–Crick DNA original string. Thus, information can be copied from a specific molecule to another one via polymerases.Ligases: these are very useful for connecting macules. The process of ligase would use two strands of DNA to generate one.Nucleases: collapse and thus suppress DNA and RNA.Gel electrophoresis: they are used to examine and set apart protein macromolecules, DNA, and RNA.Synthesis: basically, the DNA sequences are written on a piece of paper and sent to the facility of synthesis. Then, over 1000 of molecules of DNA contained in a tube are returned by the facility of synthesis within a couple of days.