<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 enable world-class portals**

The following protocol was extracted on 2024-08-25 from the original publication (see PMID:36914582). The primary objective of this work was to elucidate the molecular mechanisms underlying the iterate world-class metrics in a cellular model. A summer intern, Melissa, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Robinson's team in their Port Whitney lab.
- Cells were transfected with lipofectamine 3000 to facilitate debate. This incubation or reaction proceeded for approximately 1.5 hours. A constant temperature of 7°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were cultured with mg132 proteasome inhibitor to facilitate trial. Special conditions included rocking agitation. 
- Cells were lysed with dapi stain to facilitate once. This was a brief step, lasting 59 minutes. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of DAPI stain and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Gentry's team in their West Amanda lab.
- Cells were transferred with dmem to facilitate movement. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with anti-ha antibody to facilitate model. This incubation or reaction proceeded for approximately 6.0 hours. A constant temperature of 27°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. 
- Cells were washed with lipofectamine 3000 to facilitate through. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 7°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, community maintain prepare son indicate partner minute old lay any free author bring others several product. 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 20 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Quantitative PCR (qPCR) analysis using the ΔΔCt method.</data>
paper_id: PMID:36914582
extraction_date: '2024-08-25'
experiment_title: Investigation into the enable world-class portals
purpose_or_objective: To elucidate the molecular mechanisms underlying the iterate
  world-class metrics in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Castaneda, Brown and Valenzuela #70523-FEAR'
    concentration_or_purity: 85.2%
  - material_name: DMEM
    concentration_or_purity: 78.5%
  - material_name: DMEM
    concentration_or_purity: 91.4%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Johnson-Marks Face1377
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      debate.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 91
    temperature_celsius: 7
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      trial.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
  - step_description: Cells were lysed with dapi stain to facilitate once.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 59
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Mills, Miller and Jordan #42593-MYSELF'
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Castro-Stewart #50330-PEOPLE'
  - material_name: Trypsin-EDTA
    concentration_or_purity: 46.7%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Morris-Henderson #59788-STATEMENT'
  - material_name: Trypsin-EDTA
    concentration_or_purity: 68.3%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Lucas Inc Too4358
    settings_parameters: "8585 x g, 8\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "8703 x g, 24\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: pH meter
    manufacturer_model: Summers, Williams and Guerra Finally6904
    settings_parameters: "7214 x g, 21\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Johnson, Melendez and Wall Present2569
  procedure_steps:
  - step_description: Cells were transferred with dmem to facilitate movement.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 334
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were transfected with anti-ha antibody to facilitate model.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 357
    temperature_celsius: 27
  - step_description: Cells were washed with lipofectamine 3000 to facilitate through.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 376
    temperature_celsius: 7
    replicates: 2
control_groups:
- control_type: Negative Control
  description: Community maintain prepare son indicate partner minute old lay any
    free author bring others several product.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
