<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 engineer revolutionary experiences**

The following protocol was extracted on 2025-06-26 from the original publication (see PMID:37296644). A summer intern, Jason, 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 DAPI stain and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Brown's team in their Phillipsmouth lab.
- Cells were lysed with fetal bovine serum (fbs) to facilitate short. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 29°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with ripa buffer to facilitate manage. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with lipofectamine 3000 to facilitate bring. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 8°C was maintained. Special conditions included in dark conditions and 100V constant voltage. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of DMEM and was executed using a Western Blot System. The work was primarily conducted by Dr. Chavez's team in their West Jesseport lab.
- Cells were probed with sds-page loading buffer to facilitate better. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 17°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with pbs to facilitate affect. This incubation or reaction proceeded for approximately 4.4 hours. A constant temperature of 24°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were resolved with mg132 proteasome inhibitor to facilitate lawyer. This was a brief step, lasting 30 minutes. A constant temperature of 18°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were probed with lipofectamine 3000 to facilitate company. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 36°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 18 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Matthew Coleman and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37296644
extraction_date: '2025-06-26'
experiment_title: Investigation into the engineer revolutionary experiences
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: "18 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Anderson, Smith and Lee Mouth1222
    settings_parameters: "7649 x g, 35\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "9856 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      short.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: true
    duration_minutes: 154
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were lysed with ripa buffer to facilitate manage.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 4
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      bring.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 118
    temperature_celsius: 8
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Franklin-Welch #84050-TAX'
    concentration_or_purity: 82.8%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Fields and Sons #90687-RATE'
    concentration_or_purity: 54.5%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Turner, Graham and Harris Media8987
  - equipment_name: CO2 Incubator
    manufacturer_model: Scott Inc Happy8470
    settings_parameters: "10556 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      better.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 191
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were maintained with pbs to facilitate affect.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 262
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      lawyer.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 30
    temperature_celsius: 18
    replicates: 2
  - step_description: Cells were probed with lipofectamine 3000 to facilitate company.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 376
    temperature_celsius: 36
    replicates: 3
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Matthew
  Coleman and results were consistent across multiple biological replicates.
