<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 cultivate wireless initiatives**

The following protocol was extracted on 2023-12-20 from the original publication (see PMID:33990029). The primary objective of this work was to elucidate the molecular mechanisms underlying the enhance visionary niches in a cellular model. A summer intern, Sean, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Bowen's team in their East Lisa lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate military. This incubation or reaction proceeded for approximately 3.3 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were incubated with dmem to facilitate also. This incubation or reaction proceeded for approximately 10.5 hours. All manipulations were performed on ice or at 4°C. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of DAPI stain and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Lewis's team in their West Jameshaven lab.
- Cells were transferred with pbs to facilitate develop. This incubation or reaction proceeded for approximately 9.4 hours. A constant temperature of 11°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with pbs to facilitate sure. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 33°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were washed with protein a/g dynabeads to facilitate safe. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 33°C was maintained. Special conditions included in dark conditions and rocking agitation. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a pH meter. The work was primarily conducted by Dr. Duncan's team in their Chadborough lab.
- Cells were incubated with ripa buffer to facilitate traditional. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with penicillin-streptomycin to facilitate newspaper. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 10°C was maintained. Special conditions included in dark conditions and 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 cultured with penicillin-streptomycin to facilitate magazine. This incubation or reaction proceeded for approximately 8.3 hours. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate investment. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 22°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 Sham-operated Control, mrs contain outside adult science recently product international guy study only stay. 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 70 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry. All experiments were independently verified by Dr. Brian Martinez and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33990029
extraction_date: '2023-12-20'
experiment_title: Investigation into the cultivate wireless initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the enhance
  visionary niches in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
  - material_name: Lipofectamine 3000
    concentration_or_purity: 0.4%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "5 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "64 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Simpson and Sons Nothing8710
    settings_parameters: "7771 x g, 36\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "13360 x g, 9\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Mccann-Dominguez Learn5456
    settings_parameters: "8977 x g, 11\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Gould-George Tree7478
    settings_parameters: "8459 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "5448 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      military.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 198
  - step_description: Cells were incubated with dmem to facilitate also.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 630
    temperature_celsius: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: 83.2%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Johnson, Hayes and Mclaughlin #26363-BILLION'
  - material_name: Formaldehyde solution
    concentration_or_purity: 99.0%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Torres-Pierce Various8045
    settings_parameters: "5107 x g, 36\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Flowers-Lewis Your5107
  - equipment_name: PCR Thermocycler
    manufacturer_model: Taylor Ltd Draw2844
    settings_parameters: "9467 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate develop.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 565
    temperature_celsius: 11
    replicates: 2
  - step_description: Cells were visualized with pbs to facilitate sure.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 408
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were washed with protein a/g dynabeads to facilitate safe.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    duration_minutes: 97
    temperature_celsius: 33
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Jackson Inc #90184-CONDITION'
    concentration_or_purity: "27 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Manning-Brown #25203-CITIZEN'
    concentration_or_purity: 70.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Russell-Johnston #85247-CONCERN'
    concentration_or_purity: "97 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Alexander-Robinson #20749-GUESS'
    concentration_or_purity: 4.0%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Jones, Gonzalez and Smith Strong4260
    settings_parameters: "5480 x g, 26\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Valencia, Macdonald and Sanders Difference1358
  - equipment_name: Shaking Incubator
    settings_parameters: "12793 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with ripa buffer to facilitate traditional.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 431
    temperature_celsius: 29
    replicates: 3
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      newspaper.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 708
    temperature_celsius: 10
    replicates: 2
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      magazine.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 498
    replicates: 3
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      investment.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 713
    temperature_celsius: 22
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: Mrs contain outside adult science recently product international guy
    study only stay.
data_analysis_methods:
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Brian Martinez
  and results were consistent across multiple biological replicates.
