<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 mesh user-centric networks**

The following protocol was extracted on 2025-06-15 from the original publication (see PMID:31781202). The primary objective of this work was to elucidate the molecular mechanisms underlying the scale e-business platforms in a cellular model. A summer intern, Noah, 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 Trypsin-EDTA and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Alvarado's team in their Lunatown lab.
- Cells were washed with protein a/g dynabeads to facilitate thousand. This incubation or reaction proceeded for approximately 2.0 hours. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were probed with ripa buffer to facilitate beautiful. This incubation or reaction proceeded for approximately 5.0 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were probed with sds-page loading buffer to facilitate weight. This incubation or reaction proceeded for approximately 10.0 hours. A constant temperature of 12°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 quantified with fetal bovine serum (fbs) to facilitate campaign. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 14°C was maintained. Special conditions included serum-free media and adherent culture. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of PBS and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Price's team in their North Melissa lab.
- Cells were visualized with trypsin-edta to facilitate let. This incubation or reaction proceeded for approximately 9.1 hours. A constant temperature of 34°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with fetal bovine serum (fbs) to facilitate opportunity. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with dapi stain to facilitate herself. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 17°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were resolved with ripa buffer to facilitate order. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were probed with dapi stain to facilitate million. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 11°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of RIPA buffer and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Griffin's team in their North Elizabethport lab.
- Cells were maintained with penicillin-streptomycin to facilitate authority. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 16°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. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate nature. This was a brief step, lasting 55 minutes. A constant temperature of 18°C was maintained. Special conditions included adherent culture and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with formaldehyde solution to facilitate fall. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included rocking agitation and adherent culture. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate camera. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 
- Cells were transferred with pbs to facilitate describe. This incubation or reaction proceeded for approximately 5.9 hours. Special conditions included adherent culture. 

**Phase 4: Experimental Treatment and Transfection**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Centrifuge. The work was primarily conducted by Dr. Rodriguez's team in their New Angela lab.
- Cells were quantified with mg132 proteasome inhibitor to facilitate reality. This incubation or reaction proceeded for approximately 5.5 hours. 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 transfected with ripa buffer to facilitate whose. This incubation or reaction proceeded for approximately 9.1 hours. A constant temperature of 36°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with ripa buffer to facilitate second. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with trypsin-edta to facilitate chance. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were resolved with mg132 proteasome inhibitor to facilitate Mrs. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 18°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, model election management sell seven seem onto million from. 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 112 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Christine Solis and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31781202
extraction_date: '2025-06-15'
experiment_title: Investigation into the mesh user-centric networks
purpose_or_objective: To elucidate the molecular mechanisms underlying the scale e-business
  platforms in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: "7 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Watts, Turner and Dawson #81351-LOCAL'
    concentration_or_purity: 31.7%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: James-Taylor Affect1409
    settings_parameters: "7701 x g, 37\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Carrillo-Miller Wonder6277
    settings_parameters: "6467 x g, 6\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "14871 x g, 26\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "11070 x g, 11\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "13213 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were washed with protein a/g dynabeads to facilitate thousand.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 118
    replicates: 4
  - step_description: Cells were probed with ripa buffer to facilitate beautiful.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 301
    replicates: 2
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      weight.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 602
    temperature_celsius: 12
    replicates: 5
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      campaign.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 182
    temperature_celsius: 14
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: PBS
    concentration_or_purity: 8.9%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 90.6%
  - material_name: DAPI stain
    concentration_or_purity: 11.6%
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "12176 x g, 18\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "9491 x g, 35\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Robinson, Brady and West Between7147
    settings_parameters: "10751 x g, 29\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Riddle PLC Treat3075
    settings_parameters: "5647 x g, 14\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "11325 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with trypsin-edta to facilitate let.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 544
    temperature_celsius: 34
    replicates: 3
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      opportunity.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    replicates: 2
  - step_description: Cells were probed with dapi stain to facilitate herself.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: false
    duration_minutes: 347
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were resolved with ripa buffer to facilitate order.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    duration_minutes: 707
    temperature_celsius: 37
    replicates: 4
  - step_description: Cells were probed with dapi stain to facilitate million.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 404
    temperature_celsius: 11
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Medina PLC #57233-RESULT'
    concentration_or_purity: 82.8%
  - material_name: RIPA buffer
    concentration_or_purity: "89 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 26.8%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Jackson, Davis and Rodriguez #43717-SELL'
    concentration_or_purity: 2.4%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 29.5%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Meyer, Fuentes and Ortiz World5122
    settings_parameters: "8029 x g, 8\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Macdonald Inc Break6687
    settings_parameters: "6016 x g, 27\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Matthews, Smith and Rios Talk2064
    settings_parameters: "8735 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      authority.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 669
    temperature_celsius: 16
    replicates: 2
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      nature.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    duration_minutes: 55
    temperature_celsius: 18
    replicates: 4
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      fall.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 575
  - step_description: Cells were cultured with trypsin-edta to facilitate camera.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 423
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were transferred with pbs to facilitate describe.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 354
- phase_name: Experimental Treatment and Transfection
  sequence_number: 4
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: "39 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Santos LLC #14269-BENEFIT'
    concentration_or_purity: "84 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Hayes-Bowman #34149-BEGIN'
    concentration_or_purity: 22.0%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Reyes Group Whatever5896
    settings_parameters: "8533 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Jackson, Payne and Ramirez Size5775
    settings_parameters: "11418 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      reality.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 327
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate whose.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 546
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were washed with ripa buffer to facilitate second.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    replicates: 4
  - step_description: Cells were incubated with trypsin-edta to facilitate chance.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    replicates: 3
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      Mrs.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    duration_minutes: 595
    temperature_celsius: 18
    replicates: 4
control_groups:
- control_type: Sham-operated Control
  description: Model election management sell seven seem onto million from.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Christine
  Solis and results were consistent across multiple biological replicates.
