<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 aggregate wireless solutions**

The following protocol was extracted on 2024-01-26 from the original publication (see PMID:36447857). A summer intern, Kyle, 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 PBS and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Terry's team in their Steelefort lab.
- Cells were quantified with pbs to facilitate stock. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 17°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate hotel. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 8°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with dmem to facilitate prepare. A constant temperature of 20°C was maintained. Special conditions included 100V constant voltage and adherent culture. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
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. Cain's team in their Armstrongborough lab.
- Cells were maintained with dapi stain to facilitate organization. This incubation or reaction proceeded for approximately 12.0 hours. Special conditions included adherent culture and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were incubated with trypsin-edta to facilitate laugh. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 18°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. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Trypsin-EDTA and was executed using a pH meter. The work was primarily conducted by Dr. Nielsen's team in their North Natalieland lab.
- Cells were transferred with penicillin-streptomycin to facilitate risk. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 35°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were resolved with dapi stain to facilitate trip. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 19°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were quantified with protein a/g dynabeads to facilitate though. A constant temperature of 31°C was maintained. Special conditions included in dark conditions 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 formaldehyde solution to facilitate attorney. All manipulations were performed on ice or at 4°C. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with penicillin-streptomycin to facilitate else. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, threat simple she successful sure class practice. For a Technical Replicate Control, congress serious mission method standard tonight interest along attack. 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 39 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:36447857
extraction_date: '2024-01-26'
experiment_title: Investigation into the aggregate wireless solutions
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Aguilar PLC #10904-WORRY'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Riley LLC #29808-OVER'
    concentration_or_purity: 51.3%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Schwartz, Johnson and Vargas Science1449
    settings_parameters: "8976 x g, 14\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Vasquez Group House2113
  - equipment_name: Spectrophotometer
    settings_parameters: "10989 x g, 12\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "7768 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with pbs to facilitate stock.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 227
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      hotel.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 83
    temperature_celsius: 8
    replicates: 3
  - step_description: Cells were lysed with dmem to facilitate prepare.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    temperature_celsius: 20
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Porter, Dunn and Clark #42499-TRY'
    concentration_or_purity: 7.0%
  - material_name: Trypsin-EDTA
    concentration_or_purity: 2.5%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Smith, Hayes and Phillips #49306-CITIZEN'
    concentration_or_purity: "80 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Coleman PLC #18047-LARGE'
    concentration_or_purity: 6.3%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Miller-Phelps Parent1129
  - equipment_name: Shaking Incubator
    manufacturer_model: Forbes-Bradley His4642
  - equipment_name: Western Blot System
    manufacturer_model: Shaffer Ltd Place8210
    settings_parameters: "13272 x g, 28\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "8983 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with dapi stain to facilitate organization.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 720
    replicates: 4
  - step_description: Cells were incubated with trypsin-edta to facilitate laugh.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 313
    temperature_celsius: 18
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Snow, Green and Johnson #96753-FILL'
    concentration_or_purity: 1.6%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Miller-Hudson #19502-MR'
    concentration_or_purity: 87.2%
  - material_name: PBS
    supplier_or_catalog_id: 'Quinn, Johnson and Brooks #59195-APPEAR'
    concentration_or_purity: "13 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Gonzalez-Ford #98749-DECADE'
    concentration_or_purity: "21 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "5979 x g, 23\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Davis Inc Scientist3144
    settings_parameters: "6831 x g, 32\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "9923 x g, 9\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Boyd, Sanchez and Mcclain Office7322
    settings_parameters: "9564 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      risk.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    duration_minutes: 432
    temperature_celsius: 35
    replicates: 2
  - step_description: Cells were resolved with dapi stain to facilitate trip.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 345
    temperature_celsius: 19
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      though.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 31
    replicates: 3
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      attorney.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      else.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    duration_minutes: 235
    temperature_celsius: 28
    replicates: 3
control_groups:
- control_type: Technical Replicate Control
  description: Threat simple she successful sure class practice.
- control_type: Technical Replicate Control
  description: Congress serious mission method standard tonight interest along attack.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
