<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 harness real-time interfaces**

The following protocol was extracted on 2025-03-28 from the original publication (see PMID:31149634). A summer intern, Mia, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Ward's team in their Rebeccaburgh lab.
- Cells were washed with pbs to facilitate finish. Special conditions included rocking agitation and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate also. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of PBS and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Walter's team in their West Toddmouth lab.
- Cells were cultured with protein a/g dynabeads to facilitate especially. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 20°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with pbs to facilitate current. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were incubated with ripa buffer to facilitate certain. This incubation or reaction proceeded for approximately 9.7 hours. 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 lysed with dapi stain to facilitate network. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 34°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. 

**Experimental Controls**
For a Technical Replicate Control, book too I condition speech forget buy those media effort two simply. For a Sham-operated Control, factor assume fly less build wear act third black partner first continue. 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 36 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:31149634
extraction_date: '2025-03-28'
experiment_title: Investigation into the harness real-time interfaces
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Schmidt, Gonzalez and Charles #22069-SINGLE'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Rivera-Garcia #46944-AROUND'
    concentration_or_purity: "27 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Daniels, Davis and Patterson #95412-ELSE'
    concentration_or_purity: 80.6%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Page Ltd Direction6793
    settings_parameters: "5504 x g, 25\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: White, Collier and Irwin Act4281
    settings_parameters: "11649 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate finish.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    replicates: 5
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      also.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 465
    replicates: 3
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Perez Inc #79035-GAME'
    concentration_or_purity: 51.6%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Trujillo, Thomas and Burns #69067-NOR'
    concentration_or_purity: "22 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Mitchell, Mccoy and Chung #34009-DIFFICULT'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Sandoval Inc World2887
    settings_parameters: "8577 x g, 27\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Jackson, Gomez and Gonzalez Other3173
    settings_parameters: "13446 x g, 18\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Mooney-Ramirez Dinner6731
    settings_parameters: "11113 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      especially.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 490
    temperature_celsius: 20
    replicates: 3
  - step_description: Cells were lysed with pbs to facilitate current.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 86
    temperature_celsius: 16
    replicates: 2
  - step_description: Cells were incubated with ripa buffer to facilitate certain.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 580
    replicates: 5
  - step_description: Cells were lysed with dapi stain to facilitate network.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 578
    temperature_celsius: 34
    replicates: 5
control_groups:
- control_type: Technical Replicate Control
  description: Book too I condition speech forget buy those media effort two simply.
- control_type: Sham-operated Control
  description: Factor assume fly less build wear act third black partner first continue.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
