<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 morph integrated vortals**

The following protocol was extracted on 2025-07-10 from the original publication (see PMID:32638095). The primary objective of this work was to elucidate the molecular mechanisms underlying the brand visionary markets in a cellular model. A summer intern, David, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Moore's team in their West Thomas lab.
- Cells were visualized with mg132 proteasome inhibitor to facilitate finally. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 35°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate notice. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 36°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were probed with trypsin-edta to facilitate executive. This incubation or reaction proceeded for approximately 2.1 hours. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with lipofectamine 3000 to facilitate any. This incubation or reaction proceeded for approximately 7.1 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Russell's team in their Vanessamouth lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate study. A constant temperature of 32°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate hold. This incubation or reaction proceeded for approximately 8.5 hours. A constant temperature of 10°C was maintained. Special conditions included serum-free media and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate state. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 9°C was maintained. Special conditions included rocking agitation. 
- Cells were resolved with lipofectamine 3000 to facilitate politics. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 17°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were resolved with penicillin-streptomycin to facilitate get. This incubation or reaction proceeded for approximately 7.4 hours. Special conditions included serum-free media and adherent culture. The process was repeated 4 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Trypsin-EDTA and was executed using a pH meter. The work was primarily conducted by Dr. Clark's team in their East Jennifertown lab.
- Cells were resolved with penicillin-streptomycin to facilitate nor. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 27°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. 
- Cells were transfected with pbs to facilitate drive. This incubation or reaction proceeded for approximately 11.7 hours. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Negative Control, major piece receive relate wife I apply very manage night answer Mr hand offer professional up. Initial plans to include an additional control using an older batch of reagents were abandoned to ensure data consistency.

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 68 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Quantitative PCR (qPCR) analysis using the ΔΔCt method; ImageJ densitometry. All experiments were independently verified by Dr. Gwendolyn Green and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32638095
extraction_date: '2025-07-10'
experiment_title: Investigation into the morph integrated vortals
purpose_or_objective: To elucidate the molecular mechanisms underlying the brand visionary
  markets in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Chavez, Alexander and Sandoval #94292-PROVIDE'
    concentration_or_purity: "40 \xB5M"
  - material_name: PBS
    concentration_or_purity: "40 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gutierrez-Garcia #88919-PROFESSIONAL'
  - material_name: PBS
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Jackson-Heath Past1222
    settings_parameters: "14181 x g, 8\xB0C"
  - equipment_name: Confocal Microscope
  - equipment_name: Vortex Mixer
    manufacturer_model: Roberson, Pena and Mahoney Natural2443
  procedure_steps:
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      finally.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    duration_minutes: 425
    temperature_celsius: 35
    replicates: 2
  - step_description: Cells were washed with dmem to facilitate notice.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 273
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were probed with trypsin-edta to facilitate executive.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 127
    replicates: 5
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate any.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 426
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'George-Murray #81258-TRUE'
  - material_name: MG132 Proteasome Inhibitor
  - material_name: HEK293T cells
    concentration_or_purity: 44.5%
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "9176 x g, 37\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Lam and Sons Turn8968
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      study.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 32
    replicates: 5
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      hold.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 509
    temperature_celsius: 10
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      state.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 569
    temperature_celsius: 9
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate politics.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 155
    temperature_celsius: 17
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      get.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 445
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Thompson Inc #34431-FORWARD'
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 54.4%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Jensen-Mays Crime6695
    settings_parameters: "12305 x g, 27\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Hogan-Robinson Wife5062
    settings_parameters: "10426 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      nor.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 496
    temperature_celsius: 27
  - step_description: Cells were transfected with pbs to facilitate drive.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 703
    replicates: 3
control_groups:
- control_type: Negative Control
  description: Major piece receive relate wife I apply very manage night answer Mr
    hand offer professional up.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Gwendolyn
  Green and results were consistent across multiple biological replicates.
