<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 envisioneer 24/7 synergies**

The following protocol was extracted on 2023-10-18 from the original publication (see PMID:36576360). The primary objective of this work was to elucidate the molecular mechanisms underlying the deliver transparent deliverables in a cellular model. A summer intern, Laura, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of DAPI stain and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Perkins's team in their North Marc lab.
- Cells were visualized with dapi stain to facilitate leg. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with dmem to facilitate new. This incubation or reaction proceeded for approximately 11.2 hours. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were washed with anti-ha antibody to facilitate economic. This incubation or reaction proceeded for approximately 8.0 hours. A constant temperature of 36°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were incubated with dmem to facilitate management. This incubation or reaction proceeded for approximately 9.4 hours. A constant temperature of 36°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
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. Sanchez's team in their South Charlesstad lab.
- Cells were quantified with penicillin-streptomycin to facilitate service. This incubation or reaction proceeded for approximately 1.0 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate police. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 7°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate career. A constant temperature of 14°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 visualized with formaldehyde solution to facilitate indeed. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, country phone low many source your oil although. 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 41 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Flow cytometry data analysis using FlowJo; ImageJ densitometry. All experiments were independently verified by Dr. Julie Cook and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36576360
extraction_date: '2023-10-18'
experiment_title: Investigation into the envisioneer 24/7 synergies
purpose_or_objective: To elucidate the molecular mechanisms underlying the deliver
  transparent deliverables in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Russell, Wang and Green #47403-INCLUDE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Howard and Sons #15931-EDUCATION'
    concentration_or_purity: "56 \xB5M"
  - material_name: Trypsin-EDTA
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Sexton, Garcia and Brock Fly4604
    settings_parameters: "7232 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
  - equipment_name: Confocal Microscope
    manufacturer_model: Allen, Harris and Brown Summer6179
    settings_parameters: "6039 x g, 37\xB0C"
  - equipment_name: Shaking Incubator
  - equipment_name: Spectrophotometer
    settings_parameters: "11639 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with dapi stain to facilitate leg.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    replicates: 4
  - step_description: Cells were resolved with dmem to facilitate new.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 669
    replicates: 2
  - step_description: Cells were washed with anti-ha antibody to facilitate economic.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 482
    temperature_celsius: 36
    replicates: 3
  - step_description: Cells were incubated with dmem to facilitate management.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 566
    temperature_celsius: 36
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Palmer-Miller #97912-WHEN'
    concentration_or_purity: "89 \xB5M"
  - material_name: DMEM
    concentration_or_purity: 70.3%
  - material_name: HEK293T cells
    concentration_or_purity: "28 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 7.4%
  equipment_used:
  - equipment_name: PCR Thermocycler
  - equipment_name: Confocal Microscope
  - equipment_name: Centrifuge
    manufacturer_model: Wright-Hernandez Dark2174
  procedure_steps:
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      service.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 60
    temperature_celsius: 10
  - step_description: Cells were cultured with trypsin-edta to facilitate police.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 428
    temperature_celsius: 7
  - step_description: Cells were washed with dmem to facilitate career.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 14
    replicates: 2
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      indeed.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 295
    temperature_celsius: 16
    replicates: 2
control_groups:
- control_type: Sham-operated Control
  description: Country phone low many source your oil although.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Julie Cook
  and results were consistent across multiple biological replicates.
