Notebook

Week 1 (July 12 - July 18, 2023): Lab Training - Basic Techniques

• Day 1 (July 12): Basic Laboratory Safety Training:
  Safety protocols, lab equipment usage, and emergency procedures.
• Day 2 (July 13):: Cell Culture Training:
  Introduction to aseptic techniques, cell culture setup, and maintenance.
• Day 3 (July 14):Protein Extraction Training:
  Techniques for protein extraction and quantification.
• Day 4 (July 15):SDS-PAGE and Western Blotting Training:
  Gel electrophoresis, Western blotting setup, and antibody incubation
• Day 5 (July 16):Data Analysis Training:
  Introduction to data interpretation and analysis software.
• Day 6-7 (July 17-18):No lab procedures were performed

Week 2 (July 19 - July 25, 2023): Lab Training - Advanced Techniques

• Day 8 (July 19):Advanced Cell Culture Techniques:
  Subculturing, transfections, and specialized culture conditions.
• Day 9 (July 20):Molecular Biology Techniques:
  DNA/RNA extraction, PCR, and gene expression analysis.
• Day 10 (July 21):Advanced Western Blotting:
  Troubleshooting, quantification, and multiple antibody techniques.
• Day 11 (July 22):Microscopy and Imaging:
  Troubleshooting, quantification, and multiple antibody techniques.
• Day 12 (July 23):Biosafety and Biosecurity:
  Advanced safety procedures and regulations.
• Day 13-14 (July 24-5):No lab procedures were performed

Week 3 (July 26 - August 1 , 2023): Initial Cell Cultures

• Day 15 (July 26):Cell Culture and Establishment:
  • Prepare reagents and materials.
  • Seed cells into culture flasks/plates.
• Day 16 (July 27):Initial Culture Maintenance:
  • Check cell culture.
  • Exchange half of the medium.
• Day 17 (July 28):Ongoing Culture Maintenance. - Continue culture maintenance.
• Day 18 (July 29):Cell Harvest and Freezing:
  • Detach cells from plates.
  • Pellet cells and freeze.
• Day 19 (July 30):Cell Harvest for Protein Isolation:
  • Harvest cells for protein isolation.
  • Quantify protein concentration.
• Day 20 (July 31): Western Blotting - Block and Antibody Incubation (Day 1):
  • Prepare samples.
  • Run SDS-PAGE gel.
  • Transfer proteins to a membrane.
  • Block the membrane.
• Day 21 (August 1):Western Blotting - Washing and Antibody Incubation (Day 2):
  • Wash the membrane.
  • Incubate with primary and secondary antibodies.
  • Develop the Western blot.

Week 4 (August 2 - August 8, 2023): Cell Establishment and Early Monitoring

• Day 22 (August 2):Cell Establishment - Acquisition and Isolation:
  • Collect samples.
  • Isolate cells.
• Day 23 (August 3):Cell Establishment - Isolation of Cells:
  Isolate mononuclear cells.
• Day 24 (August 4):Cell Establishment - Isolation of Cells (contd.):
  Wash and count the isolated cells.
• Day 25 (August 5):Cell Establishment - Resuspension:
  Resuspend cells in culture medium with 20% FBS.
• Day 26 (August 6):Cell Establishment - Viability Assessment:
  Count cells and determine their viability.
• Day 27 (August 7):Cell Establishment - Storage:
  • Prepare cells for storage with DMSO.
  • Freeze and store cells for future use.
• Day 28 (August 8):Cell Proliferation and Monitoring:
  • Continue to maintain cell culture.
  • Exchange medium as needed.
  • Observe cell proliferation.
  • Cells were frozen on 80°C

---

Weeks 5-12:Experimental Continuation and Article Review

• Conduct additional literature review for updated insights.
• Incorporate new findings into the project.

Week 13: (October 4-October 10, 2023) Cells get unfrozen and cultivated,Western Blotting

• Day 85 (October 4):Cell Plating and Initial Culture:
  • Plate cells on cell culture plates.
  • Incubate at 37°C with 5% CO₂ for 48 hours (2 days).
• Day 86 (October 5):Experimental Continuation:
  Continue monitoring cell growth and experiments.
• Day 87 (October 6):Washing and Differential Medium:
  • Wash cells with PBS.
  • Divide cells into different media as per protocol.
• Day 88 (October 7):Experimental Continuation:
  Continue experiments and data collection.
• Day 89 (October 8):Cell Harvest and Freezing:
  • Harvest cells as per protocol.
  • Pellet cells and freeze for future use.
• Day 90 (October 9):Experimental Continuation:
  • Resume experiments with unfrozen cells.
  • Continue data collection.
• Day 91 (October 10):Western Blotting:
  • Block and Antibody Incubation (Day 1 of Western blotting) as per the provided protocol.
  • Begin the Western blotting process.
• Day 92 (October 11):Data Analysis and Recording:
  • Secondary Antibody Incubation (Day 2 of Western blotting)
  • Western Blotting Development
  • Analyze the data collected from the experiments.
  • Record and document the results for future reference.