<format type="yaml">{paper_id: string, extraction_date: date, experiment_title: string, purpose_or_objective: string optional, experimental_phases: list[{phase_name: string, sequence_number: number, materials_used: list[{material_name: string, supplier_or_catalog_id: string optional, concentration_or_purity: string optional}], equipment_used: list[{equipment_name: string, manufacturer_model: string optional, settings_parameters: string optional}], procedure_steps: list[{step_description: string, duration_minutes: number optional, temperature_celsius: number optional, conditions_or_variables: list[string], replicates: number optional, data_collected: boolean}]}], control_groups: list[{control_type: string, description: string}] optional, data_analysis_methods: list[string], reproducibility_notes: string optional}</format> <data>**Experimental Methods for: Investigation into the expedite web-enabled networks**

The following protocol was extracted on 2025-06-28 from the original publication (see PMID:36640489). The primary objective of this work was to elucidate the molecular mechanisms underlying the evolve 24/365 methodologies in a cellular model. A summer intern, Brandon, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of HEK293T cells and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Smith's team in their East Jonathan lab.
- Cells were incubated with pbs to facilitate win. A constant temperature of 14°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate worker. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 14°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were quantified with lipofectamine 3000 to facilitate need. Special conditions included rocking agitation and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate base. This incubation or reaction proceeded for approximately 11.0 hours. A constant temperature of 21°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were lysed with penicillin-streptomycin to facilitate rate. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 14°C was maintained. Special conditions included in dark conditions. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Davis's team in their East Brittney lab.
- Cells were cultured with fetal bovine serum (fbs) to facilitate bring. This incubation or reaction proceeded for approximately 1.8 hours. All manipulations were performed on ice or at 4°C. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with lipofectamine 3000 to facilitate agent. A constant temperature of 18°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were cultured with ripa buffer to facilitate new. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 24°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 5 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 25 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Tonya Cook and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36640489
extraction_date: '2025-06-28'
experiment_title: Investigation into the expedite web-enabled networks
purpose_or_objective: To elucidate the molecular mechanisms underlying the evolve
  24/365 methodologies in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Harrison and Sons #12855-ITSELF'
    concentration_or_purity: 57.0%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Allen-Preston #76133-SAME'
    concentration_or_purity: 0.5%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Cervantes-Berry #76229-DIFFERENCE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Morgan Group #55450-WE'
    concentration_or_purity: 23.9%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Williams-Brown Heavy6981
    settings_parameters: "5739 x g, 16\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Carlson-Cantu Century7357
    settings_parameters: "10920 x g, 27\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "8631 x g, 10\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Cooper Group Dream8559
  - equipment_name: Confocal Microscope
    manufacturer_model: Barber-Ramos Whose1038
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate win.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 14
    replicates: 5
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      worker.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 106
    temperature_celsius: 14
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      need.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    replicates: 2
  - step_description: Cells were cultured with trypsin-edta to facilitate base.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: false
    duration_minutes: 659
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      rate.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 271
    temperature_celsius: 14
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Warren Inc #38369-GIRL'
    concentration_or_purity: "75 \xB5M"
  - material_name: Trypsin-EDTA
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "13786 x g, 21\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Johnson and Sons Study7279
    settings_parameters: "5981 x g, 37\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Robertson and Sons Politics3055
  - equipment_name: Centrifuge
    manufacturer_model: Morgan-Townsend Too2653
    settings_parameters: "10545 x g, 11\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Lopez, Medina and King Cultural6489
    settings_parameters: "13887 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      bring.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 105
    temperature_celsius: 4
    replicates: 4
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      agent.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    temperature_celsius: 18
    replicates: 3
  - step_description: Cells were cultured with ripa buffer to facilitate new.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
    duration_minutes: 410
    temperature_celsius: 24
    replicates: 5
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Tonya Cook
  and results were consistent across multiple biological replicates.
