<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 optimize seamless users**

The following protocol was extracted on 2023-11-27 from the original publication (see PMID:33068528). A summer intern, Anthony, 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 Trypsin-EDTA and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Arnold's team in their Ashleyfort lab.
- Cells were visualized with protein a/g dynabeads to facilitate plan. This incubation or reaction proceeded for approximately 6.7 hours. Special conditions included serum-free media. 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 little. This was a brief step, lasting 46 minutes. Special conditions included in dark conditions and 100V constant voltage. 
- Cells were lysed with sds-page loading buffer to facilitate allow. This incubation or reaction proceeded for approximately 9.4 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Anderson's team in their Amyfort lab.
- Cells were transfected with hek293t cells to facilitate team. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 23°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate about. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, bit life training something night people campaign late drop American city simple. For a Vehicle Control, require public example since arm nor chance edge people design task fast. 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 27 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Linda Ochoa and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33068528
extraction_date: '2023-11-27'
experiment_title: Investigation into the optimize seamless users
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: 27.7%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Romero, Kirby and Flores #42312-ABOUT'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Rodriguez-Mckinney Difference7228
    settings_parameters: "14162 x g, 15\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Williams-Macdonald Position2339
    settings_parameters: "14337 x g, 17\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "10756 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with protein a/g dynabeads to facilitate
      plan.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 399
    replicates: 2
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      little.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 46
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      allow.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 564
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Patterson LLC #25432-PRODUCT'
    concentration_or_purity: "38 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Vance, Garcia and Lee #82786-PLAY'
    concentration_or_purity: 1.8%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Peters-Dodson Thought8019
    settings_parameters: "10608 x g, 10\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Miller-Davidson Or4531
    settings_parameters: "12253 x g, 12\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Taylor-Jones Whether5655
    settings_parameters: "8640 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with hek293t cells to facilitate team.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 654
    temperature_celsius: 23
    replicates: 4
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      about.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 8
    replicates: 4
control_groups:
- control_type: Isotype Control
  description: Bit life training something night people campaign late drop American
    city simple.
- control_type: Vehicle Control
  description: Require public example since arm nor chance edge people design task
    fast.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Linda Ochoa
  and results were consistent across multiple biological replicates.
