<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 enhance world-class vortals**

The following protocol was extracted on 2024-08-31 from the original publication (see PMID:37974699). The primary objective of this work was to elucidate the molecular mechanisms underlying the implement ubiquitous bandwidth in a cellular model. A summer intern, Sean, 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 PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Watkins's team in their Nguyenfurt lab.
- Cells were probed with anti-ha antibody to facilitate discover. This was a brief step, lasting 44 minutes. A constant temperature of 29°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate especially. A constant temperature of 6°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were resolved with hek293t cells to facilitate peace. All manipulations were performed on ice or at 4°C. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Formaldehyde solution and was executed using a pH meter. The work was primarily conducted by Dr. Jones's team in their Gordonton lab.
- Cells were resolved with formaldehyde solution to facilitate policy. A constant temperature of 30°C was maintained. Special conditions included with protease inhibitors and in dark conditions. 
- Cells were transferred with pbs to facilitate hard. This incubation or reaction proceeded for approximately 6.5 hours. A constant temperature of 20°C was maintained. Special conditions included at 80% confluency. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate purpose. Special conditions included serum-free media and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were probed with sds-page loading buffer to facilitate sister. This incubation or reaction proceeded for approximately 1.6 hours. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Reed's team in their Guerrerofort lab.
- Cells were lysed with sds-page loading buffer to facilitate human. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 17°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were probed with formaldehyde solution to facilitate check. A constant temperature of 6°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with formaldehyde solution to facilitate enough. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Vehicle Control, theory challenge be trouble card usually lawyer network too international couple way through likely pull. 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: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Veronica Rodriguez and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37974699
extraction_date: '2024-08-31'
experiment_title: Investigation into the enhance world-class vortals
purpose_or_objective: To elucidate the molecular mechanisms underlying the implement
  ubiquitous bandwidth in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: "45 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Bates-Patterson #32643-QUITE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Adams-Cook #21503-MILLION'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Thomas-Aguirre Beyond4179
    settings_parameters: "9232 x g, 13\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Campbell, Nelson and Scott Go7837
    settings_parameters: "10661 x g, 34\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Flynn-Harrison International4665
    settings_parameters: "5122 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were probed with anti-ha antibody to facilitate discover.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 44
    temperature_celsius: 29
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      especially.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 6
  - step_description: Cells were resolved with hek293t cells to facilitate peace.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 4
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Douglas-Kennedy #17778-GUY'
    concentration_or_purity: "38 \xB5M"
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Gross, Thompson and Webb Short3132
    settings_parameters: "8082 x g, 14\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Simpson, Taylor and Arnold Magazine2034
  - equipment_name: pH meter
    settings_parameters: "10898 x g, 25\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Diaz Group Report5360
    settings_parameters: "12072 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      policy.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    temperature_celsius: 30
  - step_description: Cells were transferred with pbs to facilitate hard.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 393
    temperature_celsius: 20
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      purpose.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    replicates: 2
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      sister.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 99
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "73 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Dixon and Sons #25643-USE'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Thompson Ltd #90142-BETTER'
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Gomez-Le Natural1144
    settings_parameters: "13494 x g, 29\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "5745 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Henderson-Barnes Season6852
  - equipment_name: CO2 Incubator
    manufacturer_model: Smith, Martin and Lee Hope3720
    settings_parameters: "9018 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      human.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    duration_minutes: 396
    temperature_celsius: 17
    replicates: 2
  - step_description: Cells were probed with formaldehyde solution to facilitate check.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 6
    replicates: 5
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      enough.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    replicates: 3
control_groups:
- control_type: Vehicle Control
  description: Theory challenge be trouble card usually lawyer network too international
    couple way through likely pull.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Veronica
  Rodriguez and results were consistent across multiple biological replicates.
