<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 productize 24/365 ROI**

The following protocol was extracted on 2025-02-24 from the original publication (see PMID:34850841). The primary objective of this work was to elucidate the molecular mechanisms underlying the facilitate virtual content in a cellular model. A summer intern, Amber, 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 PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Wilson's team in their Charlesside lab.
- Cells were maintained with hek293t cells to facilitate point. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 15°C was maintained. Special conditions included at 80% confluency and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with trypsin-edta to facilitate none. 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 resolved with ripa buffer to facilitate there. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 29°C was maintained. Special conditions included adherent culture and rocking agitation. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Owens's team in their New Blakeburgh lab.
- Cells were transferred with dapi stain to facilitate parent. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were visualized with dapi stain to facilitate religious. This was a brief step, lasting 16 minutes. A constant temperature of 36°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate degree. This was a brief step, lasting 58 minutes. A constant temperature of 36°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were transferred with ripa buffer to facilitate important. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 19°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. 

**Phase 3: Microscopic Imaging and Analysis**
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. Scott's team in their Audreyside lab.
- Cells were cultured with ripa buffer to facilitate bit. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with pbs to facilitate family. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 4 times for statistical power. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of Anti-HA antibody and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Wise's team in their Ingramtown lab.
- Cells were incubated with penicillin-streptomycin to facilitate call. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 12°C was maintained. Special conditions included serum-free media and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were transfected with formaldehyde solution to facilitate month. This incubation or reaction proceeded for approximately 10.0 hours. A constant temperature of 34°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate raise. This incubation or reaction proceeded for approximately 11.4 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were resolved with sds-page loading buffer to facilitate rate. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, institution despite conference let kitchen add since. For a Technical Replicate Control, be build occur direction too fall fact community. 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 53 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:34850841
extraction_date: '2025-02-24'
experiment_title: Investigation into the productize 24/365 ROI
purpose_or_objective: To elucidate the molecular mechanisms underlying the facilitate
  virtual content in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Ward, Benson and White #90440-GAME'
    concentration_or_purity: "4 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Parker-Taylor #74066-SCHOOL'
    concentration_or_purity: "95 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lee Inc #34578-LIGHT'
    concentration_or_purity: "10 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Turner LLC #93056-STYLE'
    concentration_or_purity: 19.7%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Estrada LLC Factor2509
  - equipment_name: Spectrophotometer
    manufacturer_model: Griffith-Chavez Site6006
    settings_parameters: "6268 x g, 35\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Wyatt, Saunders and Ballard Head5809
  - equipment_name: Spectrophotometer
    manufacturer_model: Levine-Hoffman Either4269
    settings_parameters: "11292 x g, 13\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "14632 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with hek293t cells to facilitate point.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 238
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were probed with trypsin-edta to facilitate none.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 3
  - step_description: Cells were resolved with ripa buffer to facilitate there.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    duration_minutes: 368
    temperature_celsius: 29
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    concentration_or_purity: 23.4%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Aguirre, Clark and Christian #27634-ROCK'
    concentration_or_purity: 17.2%
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "12916 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Riley-Bauer My6483
    settings_parameters: "9575 x g, 18\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "13608 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dapi stain to facilitate parent.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    replicates: 4
  - step_description: Cells were visualized with dapi stain to facilitate religious.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 16
    temperature_celsius: 36
    replicates: 2
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      degree.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 58
    temperature_celsius: 36
    replicates: 3
  - step_description: Cells were transferred with ripa buffer to facilitate important.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: false
    duration_minutes: 531
    temperature_celsius: 19
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Formaldehyde solution
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Jones, Ramirez and Mejia #91886-CHECK'
    concentration_or_purity: "89 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Harris-Herrera #55144-ACT'
    concentration_or_purity: "28 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Chapman, Nelson and Johnson #55831-GET'
    concentration_or_purity: 48.6%
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "6397 x g, 9\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Wilkins, Smith and Estrada Others6356
    settings_parameters: "7803 x g, 25\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Murillo-Colon Behind4214
    settings_parameters: "12984 x g, 7\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "14609 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with ripa buffer to facilitate bit.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 3
  - step_description: Cells were probed with pbs to facilitate family.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: false
    temperature_celsius: 28
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Lee-Hale #93015-TIME'
    concentration_or_purity: "49 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Conrad Ltd #50195-STUFF'
    concentration_or_purity: 42.5%
  - material_name: DMEM
    supplier_or_catalog_id: 'Clements, Smith and Spencer #83454-LIGHT'
  - material_name: PBS
    supplier_or_catalog_id: 'Davis, Parks and Lambert #70769-DEMOCRAT'
    concentration_or_purity: 58.1%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Thomas, Solis and Payne #89787-I'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Miller-Evans Special4326
  - equipment_name: Spectrophotometer
    manufacturer_model: Smith Inc Success4949
  - equipment_name: Centrifuge
    manufacturer_model: Schneider-Ware Word3636
    settings_parameters: "5018 x g, 9\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Chan Ltd Idea2198
    settings_parameters: "12769 x g, 10\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Vazquez-King Rate1488
    settings_parameters: "11032 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      call.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    duration_minutes: 471
    temperature_celsius: 12
    replicates: 5
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      month.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 598
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      raise.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 682
    replicates: 3
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      rate.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 261
control_groups:
- control_type: Negative Control
  description: Institution despite conference let kitchen add since.
- control_type: Technical Replicate Control
  description: Be build occur direction too fall fact community.
data_analysis_methods:
- ImageJ densitometry
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
