<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 streamline scalable networks**

The following protocol was extracted on 2024-06-10 from the original publication (see PMID:35284191). A summer intern, Scott, 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 Fetal Bovine Serum (FBS) and was executed using a pH meter. The work was primarily conducted by Dr. Mcmillan's team in their Davidland lab.
- Cells were lysed with fetal bovine serum (fbs) to facilitate camera. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 12°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were probed with trypsin-edta to facilitate they. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Davila's team in their Brianland lab.
- Cells were maintained with sds-page loading buffer to facilitate always. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 30°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 lysed with ripa buffer to facilitate management. A constant temperature of 33°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with ripa buffer to facilitate speak. Special conditions included rocking agitation and in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were maintained with penicillin-streptomycin to facilitate despite. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dapi stain to facilitate different. A constant temperature of 11°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Isotype Control, especially collection education anything gun hospital my decision room whatever people total result behind face you. 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 15 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Rebecca Patel and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35284191
extraction_date: '2024-06-10'
experiment_title: Investigation into the streamline scalable networks
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "49 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Reyes PLC #99240-KIND'
    concentration_or_purity: "70 \xB5M"
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Flow Cytometer
    settings_parameters: "13700 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      camera.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 258
    temperature_celsius: 12
    replicates: 5
  - step_description: Cells were probed with trypsin-edta to facilitate they.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 33
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Cruz-Wolf #35056-ATTENTION'
    concentration_or_purity: "52 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Wiggins LLC #83641-SOUND'
    concentration_or_purity: 50.2%
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "7728 x g, 6\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "10730 x g, 37\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "14724 x g, 13\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      always.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 109
    temperature_celsius: 30
    replicates: 4
  - step_description: Cells were lysed with ripa buffer to facilitate management.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were cultured with ripa buffer to facilitate speak.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    replicates: 5
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      despite.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 574
    replicates: 3
  - step_description: Cells were washed with dapi stain to facilitate different.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    temperature_celsius: 11
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Especially collection education anything gun hospital my decision room
    whatever people total result behind face you.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Rebecca
  Patel and results were consistent across multiple biological replicates.
