molecular techniques

• What are molecular techniques?

  Techniques to observe/manipulate molecules
• Why do we use molecular techniques?

  To study molecular make-up and functions
• What are the key components of molecular techniques?

  • Gene
  • RNA
  • Protein
  • Transcription
  • Translation
• What is the purpose of the polymerase chain reaction (PCR)?

  To amplify specific fragments of DNA
• What does the term "transcriptome" refer to?

  Dynamic expression of genes in a cell
• What is the proteome of a cell?

  Functional fingerprint of a cell at a moment
• Why do we study molecular changes in cells?

  To understand gene expression and mutations
• What is reactive gliosis?

  Hypertrophied astrocytes and scar formation
• How does exploring the molecular environment of CNS injury help?

  It aids in understanding injury repair processes
• What is the first step in assessing molecular cell make-up?

  Molecule extraction and purification
• What are the techniques used to probe the molecular make-up of a cell?

  • PCR for DNA
  • RT-PCR for RNA
  • Western blot for proteins
  • ELISA for proteins
  • Mass spectrometry for proteins
• What is the role of primers in PCR?

  To initiate DNA synthesis during amplification
• What are the three main steps of PCR?

  Denaturation, primer binding, extension
• What temperature is used for denaturation in PCR?

  95ºC
• What is the significance of primer design in PCR?

  Ensures specificity and efficiency of amplification
• How does RT-PCR differ from standard PCR?

  RT-PCR uses RNA as a template
• What is the purpose of reverse transcriptases in RT-PCR?

  To synthesize DNA from RNA templates
• What does cDNA stand for?

  Complementary DNA
• What is the application of quantitative PCR (q-PCR)?

  Real-time assessment of DNA production
• What are the differences between relative and absolute quantification in PCR?

  • Relative: Uses housekeeping gene as reference
  • Absolute: Uses known starting template DNA
• What is the significance of using a housekeeping gene in relative quantification?

  It serves as a stable reference for comparison
• What is the purpose of following cell differentiation with q-RT-PCR?

  To analyze changes in gene expression over time
• What are the applications of molecular techniques in neuroscience?

  • Gene identification
  • Mutation detection
  • Protein profiling
  • Cell differentiation analysis
  • High-throughput expression quantification
• What is the main takeaway from the summary of molecular techniques?

  PCR is a powerful tool for cell analysis
• What is the significance of the references listed in the study material?

  They provide evidence and context for techniques
• What is the focus of the neuroscience research methods discussed?

  Molecular techniques
• Why do we study proteins in neuroscience?

  To understand cell function and behavior
• What does the proteome of a cell represent?

  Functional fingerprint of the cell
• What percentage of variation in protein concentration is predicted by mRNA abundance?

  40%
• What are common methods to study proteins?

  Western blot, ELISA, Mass spectrometry
• What is the basis of the Western blot technique?

  Antibody detection
• What does ELISA stand for?

  Enzyme linked immunosorbent assay
• What is the first step in the Western blotting process?

  Protein denaturation
• What does SDS do in the Western blotting process?

  Stabilizes denatured protein and adds negative charge
• How are proteins separated in gel electrophoresis?

  Based on size
• What is the purpose of the protein transfer step in Western blotting?

  To transfer proteins to a membrane for detection
• What is used as a loading control in Western blotting?

  Β-actin
• What does PTEN/SOCS3 deletion result in regarding PI3K signaling?

  Extended PI3K signaling for longer durations
• What does the presence of growth cones indicate in nerve regeneration?

  Regenerating nerve fibers
• What are the steps involved in the Western blotting technique?

  1. Protein denaturation
  2. Addition of SDS
  3. Gel electrophoresis
  4. Protein transfer to membrane
  5. Protein detection and quantification