Molecular Biology

1. Molecular Techniques and Tools
   1. DNA Technologies
      1. Polymerase Chain Reaction (PCR)
         1. Basic principles and components
            1. DNA template
            2. Primers
            3. DNA polymerase
            4. Nucleotides
         2. Variants and applications
            1. Real-time PCR (qPCR)
            2. Reverse transcription PCR (RT-PCR)
            3. Digital PCR
         3. Troubleshooting and optimization
            1. Primer design
            2. Amplification efficiency
      2. DNA Sequencing
         1. Sanger sequencing
            1. Chain termination method
            2. Applications and limitations
         2. Next-generation sequencing (NGS)
            1. Platforms and technologies
            2. Applications in genomics and transcriptomics
         3. Third-generation sequencing
            1. Single-molecule real-time sequencing (SMRT)
            2. Nanopore sequencing
      3. Cloning and Recombinant DNA Technology
         1. Vectors and plasmids
            1. Types of vectors (e.g., plasmid, viral)
            2. Selection markers
         2. Restriction enzyme digestion
            1. Enzyme types and specificity
            2. Ligation techniques
         3. Transformation and transfection
            1. Methods: heat shock, electroporation
            2. Efficiency and selection
         4. Applications in protein production and gene therapy
   2. RNA Technologies
      1. RNA Interference (RNAi)
         1. Mechanism of action
            1. Small interfering RNA (siRNA)
            2. MicroRNA (miRNA)
         2. Applications in gene silencing and research
      2. Reverse Transcription
         1. Principles and process
         2. Reverse transcription PCR (RT-PCR)
            1. Protocols and applications
   3. Protein Analysis Techniques
      1. Western Blotting
         1. Sample preparation
         2. Gel electrophoresis
            1. SDS-PAGE
            2. Blotting methods
         3. Detection and analysis
            1. Antibody binding
            2. Detection systems (e.g., chemiluminescence)
      2. Mass Spectrometry
         1. Mass analyzers and detection
            1. Ionization techniques (e.g., MALDI, ESI)
         2. Proteomics applications
            1. Protein identification and quantification
            2. Post-translational modifications
      3. X-ray Crystallography
         1. Principles and process
         2. Crystal growth and screening
         3. Data collection and structure determination
   4. Microscopy and Imaging
      1. Fluorescence Microscopy
         1. Fluorescent dyes and proteins
         2. Imaging techniques (e.g., confocal, live-cell imaging)
         3. Applications in cell biology and diagnostics
      2. Electron Microscopy
         1. Types and techniques
            1. Transmission electron microscopy (TEM)
            2. Scanning electron microscopy (SEM)
         2. Sample preparation methods
         3. Applications in structural biology and materials science