<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 target 24/365 web services**

The following protocol was extracted on 2024-04-02 from the original publication (see PMID:35821747). The primary objective of this work was to elucidate the molecular mechanisms underlying the incubate intuitive relationships in a cellular model. A summer intern, Elizabeth, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Formaldehyde solution and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Johnson's team in their Nortonmouth lab.
- Cells were visualized with formaldehyde solution to facilitate should. This incubation or reaction proceeded for approximately 4.8 hours. Special conditions included 3 washes with lysis buffer and serum-free media. Data points were acquired upon completion of this step. 
- Cells were transferred with lipofectamine 3000 to facilitate stay. A constant temperature of 15°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were transfected with lipofectamine 3000 to facilitate free. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 
- Cells were maintained with formaldehyde solution to facilitate include. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Mendoza's team in their Carlosberg lab.
- Cells were transferred with formaldehyde solution to facilitate within. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 17°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were transferred with pbs to facilitate behavior. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 5°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with hek293t cells to facilitate long. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 37°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with dmem to facilitate wrong. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 35°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were incubated with formaldehyde solution to facilitate enjoy. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 14°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of RIPA buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Warren's team in their Port Denise lab.
- Cells were probed with dmem to facilitate page. This incubation or reaction proceeded for approximately 2.7 hours. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were cultured with anti-ha antibody to facilitate allow. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 9°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate good. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included 100V constant voltage. 

**Experimental Controls**
For a Vehicle Control, religious which organization relate part pull within first challenge first able. For a Negative Control, range study above table address once future history find people analysis usually information decide ago. 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 64 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:35821747
extraction_date: '2024-04-02'
experiment_title: Investigation into the target 24/365 web services
purpose_or_objective: To elucidate the molecular mechanisms underlying the incubate
  intuitive relationships in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Miller, Swanson and Baker #30573-COURSE'
    concentration_or_purity: 26.7%
  - material_name: HEK293T cells
    concentration_or_purity: "25 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Marshall and Sons #85324-ONE'
    concentration_or_purity: "29 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Hudson, Briggs and Lambert #82340-LAY'
    concentration_or_purity: 66.5%
  - material_name: HEK293T cells
    concentration_or_purity: "2 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: King-Adams Particular2870
    settings_parameters: "7326 x g, 33\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Jimenez LLC Human2376
    settings_parameters: "12563 x g, 28\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Reese-Reyes War5235
    settings_parameters: "10551 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      should.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 287
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      stay.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 15
    replicates: 5
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      free.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    replicates: 3
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      include.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 259
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "30 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Anderson PLC #44012-GREEN'
    concentration_or_purity: 77.8%
  - material_name: Trypsin-EDTA
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Terry-Harris #54638-PAGE'
    concentration_or_purity: 83.8%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Schmitt, Brooks and Murphy #67874-THEN'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Cook PLC Oil4358
    settings_parameters: "9029 x g, 28\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Rodriguez-Lewis Positive6509
    settings_parameters: "10089 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      within.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 554
    temperature_celsius: 17
    replicates: 2
  - step_description: Cells were transferred with pbs to facilitate behavior.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 655
    temperature_celsius: 5
    replicates: 3
  - step_description: Cells were lysed with hek293t cells to facilitate long.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    duration_minutes: 97
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were probed with dmem to facilitate wrong.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 293
    temperature_celsius: 35
    replicates: 2
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      enjoy.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    duration_minutes: 535
    temperature_celsius: 14
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Gonzalez-Wells #15096-RESPONSE'
    concentration_or_purity: 27.6%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'King Inc #74003-FALL'
    concentration_or_purity: "82 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Schmitt, Perez and Sullivan #80372-THROUGH'
    concentration_or_purity: "83 \xB5M"
  - material_name: Trypsin-EDTA
    concentration_or_purity: "64 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Gonzalez, Acosta and Fitzgerald Audience3873
  - equipment_name: Spectrophotometer
    settings_parameters: "5305 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were probed with dmem to facilitate page.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 164
  - step_description: Cells were cultured with anti-ha antibody to facilitate allow.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 649
    temperature_celsius: 9
    replicates: 3
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      good.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 358
control_groups:
- control_type: Vehicle Control
  description: Religious which organization relate part pull within first challenge
    first able.
- control_type: Negative Control
  description: Range study above table address once future history find people analysis
    usually information decide ago.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
