====== Molecular Techniques ====== Molecular techniques are laboratory methods used to study and manipulate nucleic acids (DNA, RNA) and proteins. They are essential in molecular biology, genetics, biotechnology, and clinical diagnostics. ===== 🔬 Key Techniques ===== ==== Polymerase Chain Reaction (PCR) ==== * **Purpose:** Amplifies specific DNA sequences. * **Variants:** RT-PCR (for RNA), qPCR (quantitative), multiplex PCR. * **Applications:** Pathogen detection, genotyping, cancer diagnostics. ==== Gel Electrophoresis ==== * **Purpose:** Separates DNA, RNA, or proteins based on size. * **Types:** * Agarose → for DNA/RNA. * SDS-PAGE → for proteins. * **Applications:** DNA fingerprinting, PCR product verification. ==== Nucleic Acid Extraction ==== * **Purpose:** Isolates pure DNA or RNA from biological samples. * **Applications:** PCR, sequencing, transcriptomic studies. ==== Blotting Techniques ==== * **Southern blot:** Detects specific DNA sequences. * **Northern blot:** Detects specific RNA transcripts. * **Western blot:** Detects specific proteins using antibodies. * **Applications:** Mutation analysis, gene expression, protein profiling. ==== DNA Sequencing ==== * **Sanger sequencing:** Accurate for short sequences. * **Next-Generation Sequencing (NGS):** High-throughput genome/transcriptome analysis. * **Applications:** Mutation discovery, personalized medicine, microbiome studies. ==== Microarrays ==== * **Purpose:** Analyze gene expression or detect genetic variations. * **Applications:** Transcriptomics, biomarker discovery. ==== CRISPR-Cas9 / Gene Editing ==== * **Purpose:** Precisely modify specific genes. * **Applications:** Functional genomics, gene therapy, disease modeling. ==== Flow Cytometry ==== * **Purpose:** Analyzes and sorts cells based on markers. * **Applications:** Immunophenotyping, cell cycle analysis, apoptosis. ==== ELISA (Enzyme-Linked Immunosorbent Assay) ==== * **Purpose:** Quantifies proteins, antibodies, or hormones. * **Applications:** Disease diagnosis, vaccine evaluation, immunology research. ==== RNA-Seq ==== * **Purpose:** Quantifies and sequences RNA to study gene expression. * **Applications:** Transcriptome profiling, alternative splicing, non-coding RNA detection.