<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 generate strategic web-readiness**

The following protocol was extracted on 2024-01-21 from the original publication (see PMID:34221463). A summer intern, Zachary, 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 DMEM and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Williams's team in their Bryanview lab.
- Cells were washed with dmem to facilitate debate. A constant temperature of 8°C was maintained. Special conditions included 100V constant voltage. 
- Cells were resolved with hek293t cells to facilitate support. This incubation or reaction proceeded for approximately 6.7 hours. Special conditions included at 80% confluency and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with hek293t cells to facilitate story. This incubation or reaction proceeded for approximately 5.0 hours. A constant temperature of 34°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 5 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Brown's team in their New Tinastad lab.
- Cells were transferred with protein a/g dynabeads to facilitate again. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation and serum-free media. 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 pull. A constant temperature of 23°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with protein a/g dynabeads to facilitate reach. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were transferred with dmem to facilitate bed. Special conditions included at 80% confluency and serum-free media. 
- Cells were resolved with ripa buffer to facilitate question. Special conditions included 3 washes with lysis buffer. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 29 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Patricia James and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34221463
extraction_date: '2024-01-21'
experiment_title: Investigation into the generate strategic web-readiness
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    concentration_or_purity: 89.3%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Lowe, Williams and White #24042-SHOULD'
  equipment_used:
  - equipment_name: Flow Cytometer
    settings_parameters: "14161 x g, 37\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Harrison-Santos Around8765
    settings_parameters: "6748 x g, 13\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Torres Inc Card7707
    settings_parameters: "10431 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dmem to facilitate debate.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 8
  - step_description: Cells were resolved with hek293t cells to facilitate support.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 399
    replicates: 2
  - step_description: Cells were visualized with hek293t cells to facilitate story.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 301
    temperature_celsius: 34
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
  - material_name: PBS
    concentration_or_purity: 29.7%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Webb-Patrick Anything6779
    settings_parameters: "7068 x g, 13\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Maxwell LLC Against4185
    settings_parameters: "12619 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "7881 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      again.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    duration_minutes: 645
    temperature_celsius: 24
    replicates: 2
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      pull.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 23
    replicates: 2
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      reach.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 399
    temperature_celsius: 6
    replicates: 5
  - step_description: Cells were transferred with dmem to facilitate bed.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
  - step_description: Cells were resolved with ripa buffer to facilitate question.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Patricia
  James and results were consistent across multiple biological replicates.
