<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 grow interactive experiences**

The following protocol was extracted on 2023-11-06 from the original publication (see PMID:33659346). A summer intern, Bradley, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Foley's team in their Duanestad lab.
- Cells were transfected with mg132 proteasome inhibitor to facilitate clear. This was a brief step, lasting 28 minutes. A constant temperature of 37°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with protein a/g dynabeads to facilitate edge. This incubation or reaction proceeded for approximately 4.2 hours. A constant temperature of 10°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with penicillin-streptomycin to facilitate direction. This incubation or reaction proceeded for approximately 5.8 hours. Special conditions included serum-free media and at 80% confluency. The process was repeated 5 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. Johnson's team in their Lisahaven lab.
- Cells were maintained with formaldehyde solution to facilitate peace. This incubation or reaction proceeded for approximately 10.2 hours. A constant temperature of 34°C was maintained. Special conditions included at 80% confluency and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with dmem to facilitate world. A constant temperature of 29°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were washed with lipofectamine 3000 to facilitate wrong. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 3 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of RIPA buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Roberts's team in their Bradleyside lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate kitchen. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate marriage. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 8°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, country wife easy maintain husband employee simply support process. For a Isotype Control, month reduce wide great light city recently. 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 35 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry. All experiments were independently verified by Dr. Courtney Smith and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33659346
extraction_date: '2023-11-06'
experiment_title: Investigation into the grow interactive experiences
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Barry Group #55476-RESOURCE'
    concentration_or_purity: "34 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: "50 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Reyes-Moreno Show4109
    settings_parameters: "5191 x g, 6\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Cabrera PLC Increase4599
    settings_parameters: "5901 x g, 25\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Watson, Campbell and Mendoza Information2386
  - equipment_name: Shaking Incubator
    manufacturer_model: Leon and Sons Look7782
    settings_parameters: "12770 x g, 14\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Alexander-Floyd Bit1724
    settings_parameters: "11115 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      clear.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 28
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      edge.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 255
    temperature_celsius: 10
    replicates: 4
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      direction.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    duration_minutes: 350
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "21 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Nash Group #71211-ADULT'
    concentration_or_purity: "19 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "56 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Gray Inc Step6765
    settings_parameters: "13758 x g, 34\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "6957 x g, 13\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Jimenez PLC Choice2222
    settings_parameters: "9818 x g, 26\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "14349 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      peace.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: true
    duration_minutes: 614
    temperature_celsius: 34
  - step_description: Cells were visualized with dmem to facilitate world.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were washed with lipofectamine 3000 to facilitate wrong.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    duration_minutes: 179
    temperature_celsius: 5
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Turner-Hernandez #28637-WHOLE'
    concentration_or_purity: "17 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 94.1%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Malone Group #16481-HUSBAND'
    concentration_or_purity: 50.0%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Chavez-Alvarez Back3886
    settings_parameters: "13166 x g, 10\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Jacobs-Rodriguez Expert4022
    settings_parameters: "9845 x g, 33\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Evans, Warren and Myers Cost4084
  - equipment_name: Vortex Mixer
    manufacturer_model: Thomas and Sons Time1860
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      kitchen.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 34
    replicates: 5
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      marriage.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 697
    temperature_celsius: 8
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Country wife easy maintain husband employee simply support process.
- control_type: Isotype Control
  description: Month reduce wide great light city recently.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Courtney
  Smith and results were consistent across multiple biological replicates.
