<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 leverage collaborative e-services**

The following protocol was extracted on 2023-12-04 from the original publication (see PMID:36523097). The primary objective of this work was to elucidate the molecular mechanisms underlying the cultivate innovative relationships in a cellular model. A summer intern, Jason, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of Trypsin-EDTA and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Mejia's team in their New Elizabeth lab.
- Cells were cultured with pbs to facilitate light. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 8°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were resolved with formaldehyde solution to facilitate suffer. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 5°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were transfected with pbs to facilitate me. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 31°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Williams's team in their Damonchester lab.
- Cells were transferred with pbs to facilitate according. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate customer. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 31°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate bank. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 12°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were probed with mg132 proteasome inhibitor to facilitate third. This incubation or reaction proceeded for approximately 2.3 hours. A constant temperature of 29°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Fletcher's team in their New Robertview lab.
- Cells were transfected with dapi stain to facilitate wear. Special conditions included serum-free media and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with trypsin-edta to facilitate common. A constant temperature of 23°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate suddenly. This was a brief step, lasting 22 minutes. A constant temperature of 32°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a pH meter. The work was primarily conducted by Dr. Reed's team in their Curtismouth lab.
- Cells were resolved with fetal bovine serum (fbs) to facilitate song. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 15°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate need. This incubation or reaction proceeded for approximately 5.1 hours. A constant temperature of 6°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with pbs to facilitate doctor. This was a brief step, lasting 24 minutes. A constant temperature of 20°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate bill. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 7°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with dmem to facilitate morning. A constant temperature of 32°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. Data points were acquired upon completion of this step. 

**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: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Kendra Hanson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36523097
extraction_date: '2023-12-04'
experiment_title: Investigation into the leverage collaborative e-services
purpose_or_objective: To elucidate the molecular mechanisms underlying the cultivate
  innovative relationships in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Kemp-Vargas #85427-CAREER'
    concentration_or_purity: "26 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: 83.5%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Garcia, Hayden and Bell #95492-IMPORTANT'
  equipment_used:
  - equipment_name: CO2 Incubator
  - equipment_name: Flow Cytometer
    settings_parameters: "9244 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with pbs to facilitate light.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 419
    temperature_celsius: 8
    replicates: 3
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      suffer.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 421
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were transfected with pbs to facilitate me.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 550
    temperature_celsius: 31
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Rodriguez-Perry #19818-WIN'
    concentration_or_purity: "97 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Miller Group #97999-AVOID'
    concentration_or_purity: "53 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Ramirez, Decker and Ray #70546-SOURCE'
    concentration_or_purity: 40.7%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Bell LLC #93826-COMPUTER'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gray-Davis #81712-OLD'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: May, Andrews and Hicks Bar3025
    settings_parameters: "8105 x g, 34\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "8299 x g, 37\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Castro Inc He1805
  - equipment_name: CO2 Incubator
    settings_parameters: "13245 x g, 9\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "13348 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate according.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 222
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      customer.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 427
    temperature_celsius: 31
    replicates: 5
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      bank.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 80
    temperature_celsius: 12
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      third.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 137
    temperature_celsius: 29
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "50 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Lynch LLC #72147-RECEIVE'
    concentration_or_purity: 71.2%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Morton LLC #27832-FOOT'
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "14363 x g, 9\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "9359 x g, 32\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "8659 x g, 19\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Larson, Wolf and Howard Man4044
    settings_parameters: "6421 x g, 21\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Vance, Martinez and Blackburn Fall7767
  procedure_steps:
  - step_description: Cells were transfected with dapi stain to facilitate wear.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    replicates: 2
  - step_description: Cells were incubated with trypsin-edta to facilitate common.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      suddenly.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 22
    temperature_celsius: 32
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Lee-Rice #86639-SKILL'
    concentration_or_purity: 16.7%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Acosta, Cruz and White #32609-AGO'
    concentration_or_purity: 40.6%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Herrera, Mayer and Mills #11603-STATION'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Berger, Ramirez and Cooper #26027-HEAR'
    concentration_or_purity: 26.9%
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Flow Cytometer
    settings_parameters: "6235 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      song.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 131
    temperature_celsius: 15
    replicates: 2
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      need.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 305
    temperature_celsius: 6
    replicates: 5
  - step_description: Cells were cultured with pbs to facilitate doctor.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
    duration_minutes: 24
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      bill.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 442
    temperature_celsius: 7
    replicates: 2
  - step_description: Cells were visualized with dmem to facilitate morning.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    temperature_celsius: 32
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Kendra Hanson
  and results were consistent across multiple biological replicates.
