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.