Biotechnology

  1. Core Technologies and Methodologies
    1. Recombinant DNA Technology
      1. DNA Manipulation Tools
        1. Restriction Endonucleases
          1. Type I Restriction Enzymes
            1. Type II Restriction Enzymes
              1. Type III Restriction Enzymes
                1. Recognition Sequences
                  1. Cleavage Patterns
                  2. DNA Ligases
                    1. T4 DNA Ligase
                      1. E. coli DNA Ligase
                        1. Ligation Conditions
                          1. Blunt-end vs Sticky-end Ligation
                          2. DNA Polymerases
                            1. Taq Polymerase
                              1. Pfu Polymerase
                                1. T7 DNA Polymerase
                                  1. Reverse Transcriptases
                                2. Cloning Vector Systems
                                  1. Plasmid Vectors
                                    1. pUC Series
                                      1. pBR322 Derivatives
                                        1. Expression Vectors
                                          1. Selection Markers
                                            1. Multiple Cloning Sites
                                            2. Bacteriophage Vectors
                                              1. Lambda Phage Systems
                                                1. M13 Vectors
                                                  1. Phagemid Vectors
                                                  2. Advanced Vector Systems
                                                    1. Cosmid Vectors
                                                      1. Yeast Artificial Chromosomes
                                                        1. Bacterial Artificial Chromosomes
                                                          1. P1 Artificial Chromosomes
                                                        2. Cloning Procedures
                                                          1. DNA Isolation Methods
                                                            1. Genomic DNA Extraction
                                                              1. Plasmid DNA Preparation
                                                                1. RNA Isolation Techniques
                                                                  1. DNA Purification Methods
                                                                  2. Vector Preparation
                                                                    1. Linearization Strategies
                                                                      1. Dephosphorylation
                                                                        1. Vector Purification
                                                                        2. Transformation Techniques
                                                                          1. Chemical Transformation
                                                                            1. Electroporation Methods
                                                                              1. Microinjection
                                                                                1. Biolistic Transformation
                                                                                2. Clone Selection and Screening
                                                                                  1. Antibiotic Resistance Selection
                                                                                    1. Blue-White Screening
                                                                                      1. Colony Hybridization
                                                                                        1. PCR Screening Methods
                                                                                    2. Nucleic Acid Analysis Technologies
                                                                                      1. Polymerase Chain Reaction
                                                                                        1. Standard PCR
                                                                                          1. Primer Design Principles
                                                                                            1. Thermocycling Parameters
                                                                                              1. PCR Optimization
                                                                                                1. Troubleshooting Common Problems
                                                                                                2. PCR Variations
                                                                                                  1. Real-time PCR
                                                                                                    1. Fluorescent Detection Methods
                                                                                                      1. Quantification Strategies
                                                                                                        1. Melting Curve Analysis
                                                                                                        2. Reverse Transcription PCR
                                                                                                          1. cDNA Synthesis
                                                                                                            1. One-step vs Two-step RT-PCR
                                                                                                            2. Multiplex PCR
                                                                                                              1. Primer Compatibility
                                                                                                                1. Reaction Optimization
                                                                                                                2. Long-range PCR
                                                                                                                  1. Hot-start PCR
                                                                                                                3. Electrophoretic Techniques
                                                                                                                  1. Agarose Gel Electrophoresis
                                                                                                                    1. Gel Preparation
                                                                                                                      1. Buffer Systems
                                                                                                                        1. DNA Visualization Methods
                                                                                                                          1. Size Determination
                                                                                                                          2. Polyacrylamide Gel Electrophoresis
                                                                                                                            1. Native PAGE
                                                                                                                              1. Denaturing PAGE
                                                                                                                                1. Urea-PAGE for RNA
                                                                                                                                  1. Gradient Gels
                                                                                                                                  2. Capillary Electrophoresis
                                                                                                                                    1. Principles and Applications
                                                                                                                                      1. DNA Fragment Analysis
                                                                                                                                    2. DNA Sequencing Technologies
                                                                                                                                      1. Sanger Sequencing
                                                                                                                                        1. Chain Termination Method
                                                                                                                                          1. Cycle Sequencing
                                                                                                                                            1. Automated Sequencing
                                                                                                                                              1. Data Analysis
                                                                                                                                              2. Next-Generation Sequencing
                                                                                                                                                1. Illumina Sequencing
                                                                                                                                                  1. Ion Torrent Technology
                                                                                                                                                    1. 454 Pyrosequencing
                                                                                                                                                      1. Library Preparation Methods
                                                                                                                                                        1. Data Processing Pipelines
                                                                                                                                                        2. Third-Generation Sequencing
                                                                                                                                                          1. PacBio Single-Molecule Sequencing
                                                                                                                                                            1. Oxford Nanopore Technology
                                                                                                                                                              1. Long-read Applications
                                                                                                                                                            2. Hybridization Methods
                                                                                                                                                              1. Blotting Techniques
                                                                                                                                                                1. Southern Blotting
                                                                                                                                                                  1. DNA Transfer Methods
                                                                                                                                                                    1. Probe Labeling
                                                                                                                                                                      1. Hybridization Conditions
                                                                                                                                                                      2. Northern Blotting
                                                                                                                                                                        1. RNA Transfer
                                                                                                                                                                          1. Probe Design for RNA
                                                                                                                                                                          2. Western Blotting
                                                                                                                                                                            1. Protein Transfer
                                                                                                                                                                              1. Antibody Detection
                                                                                                                                                                            2. In Situ Hybridization
                                                                                                                                                                              1. Fluorescence In Situ Hybridization
                                                                                                                                                                                1. Chromogenic Detection
                                                                                                                                                                                  1. Tissue Preparation
                                                                                                                                                                              2. Gene Editing Technologies
                                                                                                                                                                                1. CRISPR-Cas Systems
                                                                                                                                                                                  1. CRISPR-Cas9 Mechanism
                                                                                                                                                                                    1. Guide RNA Design
                                                                                                                                                                                      1. PAM Sequence Requirements
                                                                                                                                                                                        1. Cas9 Variants
                                                                                                                                                                                          1. Off-target Analysis
                                                                                                                                                                                          2. CRISPR Applications
                                                                                                                                                                                            1. Gene Knockout
                                                                                                                                                                                              1. Gene Knock-in
                                                                                                                                                                                                1. Base Editing
                                                                                                                                                                                                  1. Prime Editing
                                                                                                                                                                                                  2. Alternative CRISPR Systems
                                                                                                                                                                                                    1. CRISPR-Cas12
                                                                                                                                                                                                      1. CRISPR-Cas13
                                                                                                                                                                                                        1. dCas9 Applications
                                                                                                                                                                                                      2. Zinc Finger Nucleases
                                                                                                                                                                                                        1. Zinc Finger Protein Structure
                                                                                                                                                                                                          1. DNA Recognition Mechanisms
                                                                                                                                                                                                            1. ZFN Assembly Methods
                                                                                                                                                                                                              1. Target Site Selection
                                                                                                                                                                                                              2. TALENs Technology
                                                                                                                                                                                                                1. TALE Repeat Structure
                                                                                                                                                                                                                  1. DNA Binding Specificity
                                                                                                                                                                                                                    1. TALEN Construction
                                                                                                                                                                                                                      1. Comparison with Other Methods
                                                                                                                                                                                                                    2. Protein Technology
                                                                                                                                                                                                                      1. Protein Expression Systems
                                                                                                                                                                                                                        1. Bacterial Expression
                                                                                                                                                                                                                          1. E. coli Systems
                                                                                                                                                                                                                            1. Expression Vector Design
                                                                                                                                                                                                                              1. Inclusion Body Formation
                                                                                                                                                                                                                                1. Solubility Enhancement
                                                                                                                                                                                                                                2. Yeast Expression
                                                                                                                                                                                                                                  1. Saccharomyces cerevisiae
                                                                                                                                                                                                                                    1. Pichia pastoris
                                                                                                                                                                                                                                      1. Post-translational Modifications
                                                                                                                                                                                                                                      2. Insect Cell Expression
                                                                                                                                                                                                                                        1. Baculovirus System
                                                                                                                                                                                                                                          1. Sf9 and High Five Cells
                                                                                                                                                                                                                                            1. Protein Folding Advantages
                                                                                                                                                                                                                                            2. Mammalian Expression
                                                                                                                                                                                                                                              1. CHO Cells
                                                                                                                                                                                                                                                1. HEK293 Cells
                                                                                                                                                                                                                                                  1. Transient vs Stable Expression
                                                                                                                                                                                                                                                2. Protein Purification Strategies
                                                                                                                                                                                                                                                  1. Chromatographic Methods
                                                                                                                                                                                                                                                    1. Affinity Chromatography
                                                                                                                                                                                                                                                      1. His-tag Purification
                                                                                                                                                                                                                                                        1. GST Fusion Systems
                                                                                                                                                                                                                                                          1. Antibody Affinity
                                                                                                                                                                                                                                                          2. Ion Exchange Chromatography
                                                                                                                                                                                                                                                            1. Cation Exchange
                                                                                                                                                                                                                                                              1. Anion Exchange
                                                                                                                                                                                                                                                                1. Buffer Selection
                                                                                                                                                                                                                                                                2. Size Exclusion Chromatography
                                                                                                                                                                                                                                                                  1. Column Selection
                                                                                                                                                                                                                                                                    1. Sample Preparation
                                                                                                                                                                                                                                                                    2. Hydrophobic Interaction Chromatography
                                                                                                                                                                                                                                                                    3. Alternative Purification Methods
                                                                                                                                                                                                                                                                      1. Protein Precipitation
                                                                                                                                                                                                                                                                        1. Dialysis and Ultrafiltration
                                                                                                                                                                                                                                                                          1. Electrophoretic Purification
                                                                                                                                                                                                                                                                        2. Protein Characterization
                                                                                                                                                                                                                                                                          1. Structural Analysis
                                                                                                                                                                                                                                                                            1. X-ray Crystallography
                                                                                                                                                                                                                                                                              1. Crystal Growth
                                                                                                                                                                                                                                                                                1. Data Collection
                                                                                                                                                                                                                                                                                  1. Structure Determination
                                                                                                                                                                                                                                                                                  2. NMR Spectroscopy
                                                                                                                                                                                                                                                                                    1. Solution Structure
                                                                                                                                                                                                                                                                                      1. Protein Dynamics
                                                                                                                                                                                                                                                                                      2. Cryo-electron Microscopy
                                                                                                                                                                                                                                                                                      3. Functional Analysis
                                                                                                                                                                                                                                                                                        1. Enzyme Assays
                                                                                                                                                                                                                                                                                          1. Binding Studies
                                                                                                                                                                                                                                                                                            1. Mass Spectrometry
                                                                                                                                                                                                                                                                                              1. MALDI-TOF
                                                                                                                                                                                                                                                                                                1. ESI-MS
                                                                                                                                                                                                                                                                                                  1. Protein Identification
                                                                                                                                                                                                                                                                                            2. Cell and Tissue Culture
                                                                                                                                                                                                                                                                                              1. Sterile Technique
                                                                                                                                                                                                                                                                                                1. Aseptic Practices
                                                                                                                                                                                                                                                                                                  1. Laminar Flow Hoods
                                                                                                                                                                                                                                                                                                    1. Sterilization Methods
                                                                                                                                                                                                                                                                                                      1. Contamination Detection
                                                                                                                                                                                                                                                                                                      2. Microbial Culture Systems
                                                                                                                                                                                                                                                                                                        1. Batch Culture
                                                                                                                                                                                                                                                                                                          1. Continuous Culture
                                                                                                                                                                                                                                                                                                            1. Fed-batch Culture
                                                                                                                                                                                                                                                                                                              1. Bioreactor Design
                                                                                                                                                                                                                                                                                                              2. Animal Cell Culture
                                                                                                                                                                                                                                                                                                                1. Cell Line Establishment
                                                                                                                                                                                                                                                                                                                  1. Primary Cell Isolation
                                                                                                                                                                                                                                                                                                                    1. Immortalization Methods
                                                                                                                                                                                                                                                                                                                      1. Cell Line Characterization
                                                                                                                                                                                                                                                                                                                      2. Culture Conditions
                                                                                                                                                                                                                                                                                                                        1. Media Formulation
                                                                                                                                                                                                                                                                                                                          1. Serum Requirements
                                                                                                                                                                                                                                                                                                                            1. Growth Factor Supplementation
                                                                                                                                                                                                                                                                                                                              1. pH and Gas Control
                                                                                                                                                                                                                                                                                                                              2. Cell Preservation
                                                                                                                                                                                                                                                                                                                                1. Cryopreservation Protocols
                                                                                                                                                                                                                                                                                                                                  1. Freezing Media
                                                                                                                                                                                                                                                                                                                                    1. Thawing Procedures
                                                                                                                                                                                                                                                                                                                                  2. Plant Tissue Culture
                                                                                                                                                                                                                                                                                                                                    1. Explant Preparation
                                                                                                                                                                                                                                                                                                                                      1. Callus Induction
                                                                                                                                                                                                                                                                                                                                        1. Shoot Regeneration
                                                                                                                                                                                                                                                                                                                                          1. Root Development
                                                                                                                                                                                                                                                                                                                                            1. Acclimatization

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                                                                                                                                                                                                                                                                                                                                        1. Foundations of Biotechnology

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                                                                                                                                                                                                                                                                                                                                        3. Applications of Biotechnology