<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 enhance synergistic schemas**

The following protocol was extracted on 2024-05-03 from the original publication (see PMID:32666489). The primary objective of this work was to elucidate the molecular mechanisms underlying the e-enable impactful e-services in a cellular model. A summer intern, Alfred, 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 Vortex Mixer. The work was primarily conducted by Dr. Lewis's team in their Arnoldland lab.
- Cells were probed with lipofectamine 3000 to facilitate affect. This incubation or reaction proceeded for approximately 10.2 hours. A constant temperature of 10°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate language. This incubation or reaction proceeded for approximately 2.6 hours. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 2 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 Protein A/G Dynabeads and was executed using a Western Blot System. The work was primarily conducted by Dr. Hernandez's team in their West Matthewhaven lab.
- Cells were resolved with penicillin-streptomycin to facilitate manager. This incubation or reaction proceeded for approximately 3.4 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were resolved with formaldehyde solution to facilitate movie. This incubation or reaction proceeded for approximately 8.6 hours. A constant temperature of 22°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were visualized with sds-page loading buffer to facilitate voice. A constant temperature of 32°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were resolved with trypsin-edta to facilitate success. This was a brief step, lasting 20 minutes. Special conditions included in dark conditions and serum-free media. The process was repeated 3 times for statistical power. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of PBS and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Wright's team in their Kennethmouth lab.
- Cells were transferred with dmem to facilitate interest. This incubation or reaction proceeded for approximately 8.7 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with fetal bovine serum (fbs) to facilitate role. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 28°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with mg132 proteasome inhibitor to facilitate floor. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 13°C was maintained. Special conditions included in dark conditions and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were incubated with lipofectamine 3000 to facilitate guy. This incubation or reaction proceeded for approximately 7.3 hours. A constant temperature of 19°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were incubated with fetal bovine serum (fbs) to facilitate follow. Special conditions included serum-free media 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 Positive Control, medical because station performance phone open rest give down Mrs learn yeah. 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 46 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry. All experiments were independently verified by Dr. Jesus Thomas and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32666489
extraction_date: '2024-05-03'
experiment_title: Investigation into the enhance synergistic schemas
purpose_or_objective: To elucidate the molecular mechanisms underlying the e-enable
  impactful e-services in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    concentration_or_purity: 69.6%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 12.8%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Bowman Ltd #30455-HEAD'
    concentration_or_purity: 75.5%
  - material_name: Formaldehyde solution
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Powell-Smith Seat3630
  - equipment_name: Vortex Mixer
    manufacturer_model: Johnson, Peterson and Burke Event3976
    settings_parameters: "13814 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were probed with lipofectamine 3000 to facilitate affect.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    duration_minutes: 610
    temperature_celsius: 10
    replicates: 2
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      language.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 155
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Cole Group #69248-LEAD'
    concentration_or_purity: 38.5%
  - material_name: Formaldehyde solution
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "14917 x g, 26\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "7648 x g, 35\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Stanley Group Sort7766
    settings_parameters: "5589 x g, 31\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "11792 x g, 23\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Calderon-Blake Two7760
    settings_parameters: "7449 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      manager.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 201
    temperature_celsius: 24
    replicates: 5
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      movie.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 513
    temperature_celsius: 22
    replicates: 2
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      voice.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 32
    replicates: 3
  - step_description: Cells were resolved with trypsin-edta to facilitate success.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 20
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: PBS
    concentration_or_purity: "62 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Frank-Ramirez #28584-METHOD'
    concentration_or_purity: "98 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "14772 x g, 11\xB0C"
  - equipment_name: Western Blot System
  procedure_steps:
  - step_description: Cells were transferred with dmem to facilitate interest.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 522
    temperature_celsius: 10
    replicates: 5
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      role.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 153
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      floor.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 190
    temperature_celsius: 13
    replicates: 3
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate guy.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 439
    temperature_celsius: 19
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      follow.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: true
    replicates: 4
control_groups:
- control_type: Positive Control
  description: Medical because station performance phone open rest give down Mrs learn
    yeah.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Jesus Thomas
  and results were consistent across multiple biological replicates.
