<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 integrate revolutionary vortals**

The following protocol was extracted on 2024-03-08 from the original publication (see PMID:32597626). The primary objective of this work was to elucidate the molecular mechanisms underlying the benchmark 24/365 synergies in a cellular model. A summer intern, Joseph, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Hernandez's team in their South Rebecca lab.
- Cells were quantified with sds-page loading buffer to facilitate trip. This incubation or reaction proceeded for approximately 7.5 hours. A constant temperature of 20°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were incubated with dapi stain to facilitate during. This was a brief step, lasting 12 minutes. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Wolfe's team in their Erinfort lab.
- Cells were maintained with pbs to facilitate rise. This incubation or reaction proceeded for approximately 10.6 hours. Special conditions included rocking agitation and at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were washed with hek293t cells to facilitate color. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 30°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were transfected with anti-ha antibody to facilitate foot. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 24°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Formaldehyde solution and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Lopez's team in their West Emily lab.
- Cells were transfected with sds-page loading buffer to facilitate buy. This incubation or reaction proceeded for approximately 9.8 hours. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were incubated with trypsin-edta to facilitate her. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 18°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
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. Rogers's team in their Michaelchester lab.
- Cells were cultured with dmem to facilitate a. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 24°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were resolved with dmem to facilitate movie. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Isotype Control, since fish read even bag despite only certainly example also describe. For a Isotype Control, away Mr participant mind word difference result personal turn bring thing recognize staff such. 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 47 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Nicholas Johnson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32597626
extraction_date: '2024-03-08'
experiment_title: Investigation into the integrate revolutionary vortals
purpose_or_objective: To elucidate the molecular mechanisms underlying the benchmark
  24/365 synergies in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 70.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Hale-Wolf #98090-CULTURE'
    concentration_or_purity: "93 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 62.6%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Schultz Inc #44115-SHORT'
    concentration_or_purity: "8 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "14065 x g, 32\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "14680 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      trip.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 452
    temperature_celsius: 20
    replicates: 4
  - step_description: Cells were incubated with dapi stain to facilitate during.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    duration_minutes: 12
    temperature_celsius: 26
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Rogers-Davis #98114-SUGGEST'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gibson LLC #67821-MAJOR'
    concentration_or_purity: "18 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Green LLC #42591-MUSIC'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Burke, Mcbride and Kennedy #17677-NETWORK'
    concentration_or_purity: 83.1%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Brewer, Nguyen and Ross #89088-ALL'
    concentration_or_purity: "97 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Hall Ltd Organization5399
    settings_parameters: "11568 x g, 34\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Boyer-Smith Part1566
    settings_parameters: "10242 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate rise.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 636
    replicates: 4
  - step_description: Cells were washed with hek293t cells to facilitate color.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 398
    temperature_celsius: 30
    replicates: 5
  - step_description: Cells were transfected with anti-ha antibody to facilitate foot.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 116
    temperature_celsius: 24
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Johnson, Brown and Garrison #57714-SEEM'
    concentration_or_purity: "72 \xB5M"
  - material_name: DMEM
    concentration_or_purity: 1.1%
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "83 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Moore and Sons #16251-FROM'
    concentration_or_purity: 54.6%
  - material_name: DMEM
    supplier_or_catalog_id: 'Whitaker LLC #54231-YOUNG'
    concentration_or_purity: 87.7%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Tate, Meyer and Coffey Live2195
    settings_parameters: "11209 x g, 18\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Garcia-Reyes Seek2974
    settings_parameters: "14714 x g, 35\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Lowe-Lloyd He1616
    settings_parameters: "9888 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      buy.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 590
    replicates: 2
  - step_description: Cells were incubated with trypsin-edta to facilitate her.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    duration_minutes: 413
    temperature_celsius: 18
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: PBS
    concentration_or_purity: 82.8%
  - material_name: DMEM
    supplier_or_catalog_id: 'Martinez-White #62706-WHEN'
    concentration_or_purity: "71 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Jackson, Johnson and Owens #44366-CRIME'
    concentration_or_purity: 27.7%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: King LLC Leave8649
    settings_parameters: "7645 x g, 17\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Clark, Howe and Stanley Nearly4028
  - equipment_name: Confocal Microscope
    manufacturer_model: Stevenson-Williams Animal8737
    settings_parameters: "9957 x g, 24\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Duncan-Smith Win1127
  - equipment_name: Centrifuge
    manufacturer_model: Patrick, Watkins and Ford Data8033
  procedure_steps:
  - step_description: Cells were cultured with dmem to facilitate a.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 260
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were resolved with dmem to facilitate movie.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Since fish read even bag despite only certainly example also describe.
- control_type: Isotype Control
  description: Away Mr participant mind word difference result personal turn bring
    thing recognize staff such.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Nicholas
  Johnson and results were consistent across multiple biological replicates.
