Molecular Techniques [Neurosurgery Wiki]

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====== 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.