BACE Exam-Knowledge Question and Answer 2023

Clinical Trial- Phase 1 - - Tests drug in healthy individuals - Tests for safety, dosage, and side effects Clinical Trial- Phase 2 - - Tests on larger group of effected individuals - Tests for efficacy and side effects Clinical Trial-Phase 3 - - Tests on new and wider demographic - Tests for long term effectiveness and comparisons with other medications Clinical Trial- FDA Approval - Treatment determined effective and safe for public use Clinical Trial- Phase 4 - - Continues to test for effectiveness and safety - Can be taken off the market if necessary Cell Harvesting- Indirect Cell Count - Determine concentration of cells is a liquid culture DOES NOT tell me if the cells are viable vs nonviable- some cells may be dead Cell Harvesting- Indirect Cell Count Reagents - - Luria Broth(Food Source) - Shaking Incubator or shaking water bath (Incubator or Water is for 37 C; Shaking-Oxygen) - Spectrophotometer (measures absorbance values to give concentration) Cell Harvesting- Direct Cell Count - Determine concentration of cells in agar place Cell Harvesting- Direct Cell Count Reagents - - Agar Plate - Luria Broth(food source) - Incubator (37 C) - Physically Count- the amount the colonies and calculate the concentration Gram Staining - Determine if peptidoglycan is present in cell wall Gram Staining Reagents - - Crystal Violet=Gram Positive - Iodine= fixation - Alcohol= decolorizer - Safranin= Gram negative Inserting into the Cell - Neutralize cell membrane and substance being inserted now passive disuse can occur Inserting into the Cell- Reagents - - Cofactors: CaCl2 or MgCl2 - Heat-shock (chemical transformation= prokaryotic) or Electricity/Electroporation (Transfection= Eukaryotic Cells) Lysing the Cell - Breaking (destroying cell membrane) open the cell to extract DNA, mRNA, or protein Lysing the Cell- Reagents - - Detergent: SDS (DNA Extraction) or Lysozyme (Protein Extraction) - Freeze-Thaw Method: Freezer, water bath/dry bath, vortexer PCR - Copy DNA PCR- Reagents - - DNA Sample (denaturing) - Primers (annealing) - Nucleotides/dNTPs (elongation) - Taq Polymerase (elongation) - Mix Buffer (maintain pH) - PCR tube