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Lab Notebook

Week 1: Inspiration from other teams
Week 2: Delivery of plasmid to prostate
Week 3: Guides of polymers - Aptamers
Week 1: Formulation of initial plan
  1. Construction of plasmids
  2. Purchase of A and B separately
  1. A plasmid containing our gene that is non-mammalian; and
  2. Another plasmid that is mammalian but lacks any promoter genes (pENTR1A)
    1. Testing of PSMA promoter
      • Evaluate the activity of PSMA promoter in different cell lines
      • GFP (commonly used and known to work) as reporter gene
      • Transfected to cancer and non-cancerous cell lines
    1. Testing of Gluc reporter gene
      • Measure luminescence given out by reaction w/ Gluc as substrate
      • Transfected to cancer and non-cancerous cell lines
      • Testing of polymer delivery
      • Make sure polymers can correctly transport plasmids to cancer cells
    1. Testing of killing function
      • Confirm that Bax gene can kill cancer cells
Week 2: Figuring out protocols used
  1. Plasmid construction
    1. Plasmids in powder form: transformed into DH5a
    2. Colonies are picked → plasmids extracted
    3. Restriction digestion: linearise genes (BamHI and HindIII)
    4. Gel electrophoresis: visualise the results of digestion
    5. Gel purification: obtain the digested plasmids
    6. Ligation: stick two genes together
    7. Colony PCR: confirm that ligated genes have the correct size
  2. Polymers
    1. PEG is attached to PAMAM to form PAMAM-PEG conjugate
    2. Aptamers are attached to PAMAM-PEG
    3. BODIPY are introduced to some of the PAMAM-PEG
      1. Microscopes are used to view BODIPY fluorescence
      2. Confirm that aptamers can bring plasmids to cells
    4. Plasmids C & D are introduced to other PAMAM-PEG
    5. Polymer-plasmid conjugates are mixed with cells
      1. MTT Assay are performed to quantify the death of cancer cells (required absorbance measurement)
      2. Luminescence of Gluc are detected
Week 3: Required equipment and apparatus
Week 4: Evaluation of initial plan
July-September Journal

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Game Development Notebook

March
April
May-June
July-September
Technical Issues
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