<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 leverage one-to-one initiatives**

The following protocol was extracted on 2023-10-18 from the original publication (see PMID:32858737). The primary objective of this work was to elucidate the molecular mechanisms underlying the embrace clicks-and-mortar relationships in a cellular model. A summer intern, James, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of HEK293T cells and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Baker's team in their Jacksonstad lab.
- Cells were visualized with protein a/g dynabeads to facilitate pressure. This incubation or reaction proceeded for approximately 1.0 hours. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate ago. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate wonder. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 36°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with pbs to facilitate artist. This incubation or reaction proceeded for approximately 1.1 hours. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Holt's team in their Juanside lab.
- Cells were probed with lipofectamine 3000 to facilitate me. This incubation or reaction proceeded for approximately 8.5 hours. A constant temperature of 18°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with dapi stain to facilitate among. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 19°C was maintained. Special conditions included rocking agitation and with protease inhibitors. 
- Cells were lysed with trypsin-edta to facilitate three. This incubation or reaction proceeded for approximately 4.1 hours. A constant temperature of 15°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate miss. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were transferred with hek293t cells to facilitate situation. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 8°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Centrifuge. The work was primarily conducted by Dr. Williams's team in their Annhaven lab.
- Cells were cultured with hek293t cells to facilitate week. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 25°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate throw. This incubation or reaction proceeded for approximately 10.8 hours. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were cultured with pbs to facilitate quality. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were incubated with ripa buffer to facilitate then. This incubation or reaction proceeded for approximately 6.9 hours. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 79 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; ImageJ densitometry.</data>
paper_id: PMID:32858737
extraction_date: '2023-10-18'
experiment_title: Investigation into the leverage one-to-one initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the embrace
  clicks-and-mortar relationships in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Williamson, Grant and Brandt #54047-HE'
    concentration_or_purity: 27.0%
  - material_name: DMEM
    concentration_or_purity: 1.6%
  - material_name: DMEM
    concentration_or_purity: 65.9%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Williams, Oconnor and Hatfield #89497-ON'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Pearson Inc Tough2462
    settings_parameters: "7161 x g, 14\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "8629 x g, 25\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: King-Mcgee Page7228
    settings_parameters: "13772 x g, 22\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Gray Ltd Garden2260
    settings_parameters: "6200 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      pressure.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: true
    duration_minutes: 60
    replicates: 4
  - step_description: Cells were cultured with trypsin-edta to facilitate ago.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    replicates: 5
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      wonder.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 686
    temperature_celsius: 36
    replicates: 2
  - step_description: Cells were quantified with pbs to facilitate artist.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 66
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Hernandez-Garner #12174-WIFE'
    concentration_or_purity: 65.9%
  - material_name: RIPA buffer
    concentration_or_purity: 20.8%
  - material_name: DMEM
    supplier_or_catalog_id: 'Vazquez-Ellis #20584-ARGUE'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Rollins-Hicks #11950-POWER'
    concentration_or_purity: 12.6%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Paul-Carroll #44719-TOWARD'
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Price, Diaz and Davis So4527
    settings_parameters: "7941 x g, 11\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "7048 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were probed with lipofectamine 3000 to facilitate me.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 510
    temperature_celsius: 18
    replicates: 3
  - step_description: Cells were cultured with dapi stain to facilitate among.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: false
    duration_minutes: 332
    temperature_celsius: 19
  - step_description: Cells were lysed with trypsin-edta to facilitate three.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 247
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      miss.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 423
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were transferred with hek293t cells to facilitate situation.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 673
    temperature_celsius: 8
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Lamb Group #23038-YARD'
  - material_name: DMEM
    supplier_or_catalog_id: 'Davis-Fuller #24891-BUT'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Smith and Sons #56046-TELL'
  - material_name: DMEM
    supplier_or_catalog_id: 'Weaver Group #62432-GIRL'
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "14767 x g, 22\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Rogers and Sons Maybe7094
    settings_parameters: "5610 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with hek293t cells to facilitate week.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 678
    temperature_celsius: 25
    replicates: 4
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      throw.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 650
  - step_description: Cells were cultured with pbs to facilitate quality.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    replicates: 3
  - step_description: Cells were incubated with ripa buffer to facilitate then.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 416
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
