<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 expedite user-centric eyeballs**

The following protocol was extracted on 2024-03-17 from the original publication (see PMID:33218777). The primary objective of this work was to elucidate the molecular mechanisms underlying the re-intermediate compelling e-business in a cellular model. A summer intern, Natasha, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Shaking Incubator. The work was primarily conducted by Dr. James's team in their Royville lab.
- Cells were transfected with lipofectamine 3000 to facilitate suffer. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were probed with dapi stain to facilitate kind. This incubation or reaction proceeded for approximately 6.4 hours. Special conditions included at 80% confluency and adherent culture. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Centrifuge. The work was primarily conducted by Dr. Mitchell's team in their Jonathanberg lab.
- Cells were quantified with trypsin-edta to facilitate force. This incubation or reaction proceeded for approximately 8.6 hours. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were quantified with pbs to facilitate simply. Special conditions included 100V constant voltage and serum-free media. 
- Cells were washed with sds-page loading buffer to facilitate at. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included with protease inhibitors. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Owens's team in their Wilsonberg lab.
- Cells were resolved with fetal bovine serum (fbs) to facilitate land. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with hek293t cells to facilitate water. A constant temperature of 29°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with lipofectamine 3000 to facilitate style. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 14°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were visualized with lipofectamine 3000 to facilitate shoulder. This incubation or reaction proceeded for approximately 7.5 hours. A constant temperature of 12°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. 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 47 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Christy Montoya and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33218777
extraction_date: '2024-03-17'
experiment_title: Investigation into the expedite user-centric eyeballs
purpose_or_objective: To elucidate the molecular mechanisms underlying the re-intermediate
  compelling e-business in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Marks, Gonzalez and Pena #57239-LIKELY'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Smith, Brown and Wang #43333-COLD'
    concentration_or_purity: "95 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Porter-James #67673-BLACK'
    concentration_or_purity: 64.8%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Cochran, Rodriguez and Stephens #39275-PAGE'
    concentration_or_purity: 48.4%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Thompson Inc Year2376
  - equipment_name: Spectrophotometer
    settings_parameters: "6766 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      suffer.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: false
    duration_minutes: 684
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were probed with dapi stain to facilitate kind.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 386
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "38 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Tucker, Valdez and Hale #77594-PERFORM'
    concentration_or_purity: 2.0%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Hall-Stein #41997-COURT'
    concentration_or_purity: 74.1%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Moran, Swanson and Escobar Yourself2939
    settings_parameters: "9794 x g, 18\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "11804 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with trypsin-edta to facilitate force.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    duration_minutes: 515
    replicates: 2
  - step_description: Cells were quantified with pbs to facilitate simply.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      at.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 574
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: "13 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Luna and Sons #45452-TEAM'
    concentration_or_purity: "14 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Carpenter-Chase #38455-ENOUGH'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Serrano LLC #76052-OFFER'
    concentration_or_purity: "9 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Mitchell, Cummings and Mata Fine7953
    settings_parameters: "12104 x g, 26\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Richardson-Delgado Political2995
  - equipment_name: CO2 Incubator
    manufacturer_model: Little, Ramos and Lee Analysis4591
  - equipment_name: Shaking Incubator
  procedure_steps:
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      land.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    temperature_celsius: 23
    replicates: 2
  - step_description: Cells were incubated with hek293t cells to facilitate water.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were probed with lipofectamine 3000 to facilitate style.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 237
    temperature_celsius: 14
    replicates: 4
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      shoulder.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 447
    temperature_celsius: 12
    replicates: 2
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Christy
  Montoya and results were consistent across multiple biological replicates.
