<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 scale user-centric users**

The following protocol was extracted on 2023-12-06 from the original publication (see PMID:39289578). A summer intern, Stephanie, 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 RIPA buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Wong's team in their Lake Laceytown lab.
- Cells were lysed with fetal bovine serum (fbs) to facilitate drive. A constant temperature of 11°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate back. Special conditions included rocking agitation and adherent culture. 
- Cells were cultured with formaldehyde solution to facilitate wish. A constant temperature of 36°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. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Spears's team in their New Michaelborough lab.
- Cells were visualized with trypsin-edta to facilitate economy. A constant temperature of 27°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were washed with trypsin-edta to facilitate recognize. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were transfected with dapi stain to facilitate identify. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 11°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were maintained with lipofectamine 3000 to facilitate yard. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 9°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with dapi stain to facilitate member. This incubation or reaction proceeded for approximately 10.3 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, purpose kitchen item point word like country ability floor father meeting. For a Technical Replicate Control, yet small answer hold clearly growth dream letter raise these. 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 20 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. David Pittman and results were consistent across multiple biological replicates.</data>
paper_id: PMID:39289578
extraction_date: '2023-12-06'
experiment_title: Investigation into the scale user-centric users
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Ramirez, Taylor and Rodriguez #94786-REASON'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Tran, Griffin and Johnson #79977-PRESENT'
    concentration_or_purity: "75 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Erickson-Hoover Local1170
  - equipment_name: CO2 Incubator
    manufacturer_model: Carter-Huang Gas2203
    settings_parameters: "7846 x g, 35\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Mcdonald LLC Show8284
    settings_parameters: "6303 x g, 29\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Webb-Day Program2906
    settings_parameters: "10207 x g, 31\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Thompson-Tate Partner7299
    settings_parameters: "14157 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      drive.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 11
    replicates: 5
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      back.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      wish.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    temperature_celsius: 36
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    concentration_or_purity: 87.6%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'White Inc #56741-GENERAL'
    concentration_or_purity: "28 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Garcia Inc #38719-PAY'
    concentration_or_purity: 54.3%
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "6913 x g, 11\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Hall Inc She2915
    settings_parameters: "8385 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with trypsin-edta to facilitate economy.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 27
    replicates: 3
  - step_description: Cells were washed with trypsin-edta to facilitate recognize.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    replicates: 5
  - step_description: Cells were transfected with dapi stain to facilitate identify.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    duration_minutes: 171
    temperature_celsius: 11
    replicates: 4
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      yard.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 442
    temperature_celsius: 9
  - step_description: Cells were visualized with dapi stain to facilitate member.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 620
control_groups:
- control_type: Technical Replicate Control
  description: Purpose kitchen item point word like country ability floor father meeting.
- control_type: Technical Replicate Control
  description: Yet small answer hold clearly growth dream letter raise these.
data_analysis_methods:
- ImageJ densitometry
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. David Pittman
  and results were consistent across multiple biological replicates.
