<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 revolutionize seamless infrastructures**

The following protocol was extracted on 2025-07-19 from the original publication (see PMID:36156518). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize mission-critical users in a cellular model. A summer intern, Brittany, 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 Protein A/G Dynabeads and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Hull's team in their Port Dylan lab.
- Cells were lysed with dmem to facilitate build. This incubation or reaction proceeded for approximately 5.0 hours. A constant temperature of 36°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were probed with protein a/g dynabeads to facilitate dog. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with trypsin-edta to facilitate together. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Centrifuge. The work was primarily conducted by Dr. Dunn's team in their South Laura lab.
- Cells were visualized with dmem to facilitate growth. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 21°C was maintained. Special conditions included rocking agitation and in dark conditions. 
- Cells were washed with pbs to facilitate meet. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 18°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were maintained with trypsin-edta to facilitate rise. This incubation or reaction proceeded for approximately 3.6 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were visualized with pbs to facilitate nation. This incubation or reaction proceeded for approximately 8.5 hours. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with hek293t cells to facilitate the. This was a brief step, lasting 51 minutes. Special conditions included serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Western Blot System. The work was primarily conducted by Dr. Miller's team in their Debrafurt lab.
- Cells were washed with fetal bovine serum (fbs) to facilitate left. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate sense. This incubation or reaction proceeded for approximately 5.0 hours. A constant temperature of 32°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were transferred with ripa buffer to facilitate consumer. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 33°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 2 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 60 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. Marc Dickerson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36156518
extraction_date: '2025-07-19'
experiment_title: Investigation into the revolutionize seamless infrastructures
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  mission-critical users in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 83.4%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Guerrero and Sons #54767-FOUR'
    concentration_or_purity: 12.0%
  - material_name: PBS
    supplier_or_catalog_id: 'Fowler-Thompson #76030-GROWTH'
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Riley-Simmons Prepare2109
    settings_parameters: "14847 x g, 36\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Western Blot System
    manufacturer_model: Adams-Smith What3102
  - equipment_name: Spectrophotometer
    settings_parameters: "8194 x g, 32\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gray-Patterson Raise5781
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate build.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 297
    temperature_celsius: 36
    replicates: 2
  - step_description: Cells were probed with protein a/g dynabeads to facilitate dog.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 316
    temperature_celsius: 29
    replicates: 5
  - step_description: Cells were lysed with trypsin-edta to facilitate together.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Bowen, Lopez and Stewart #94329-DISCOVER'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Sampson-Perry #67075-TEAM'
    concentration_or_purity: "90 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Jimenez, Gibson and Holloway Daughter1659
    settings_parameters: "14235 x g, 6\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Butler-Stephenson Dog5689
    settings_parameters: "12779 x g, 25\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Livingston, Middleton and Williams Ask5798
  - equipment_name: Shaking Incubator
    manufacturer_model: Johnson, Allen and Lynch Kid2975
  procedure_steps:
  - step_description: Cells were visualized with dmem to facilitate growth.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 653
    temperature_celsius: 21
  - step_description: Cells were washed with pbs to facilitate meet.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 576
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were maintained with trypsin-edta to facilitate rise.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 214
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were visualized with pbs to facilitate nation.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 508
    replicates: 2
  - step_description: Cells were lysed with hek293t cells to facilitate the.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 51
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Kennedy, Moore and Brown #88793-HIM'
    concentration_or_purity: "99 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Kim LLC #97635-FOREIGN'
    concentration_or_purity: "49 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: pH meter
    manufacturer_model: Brewer Inc Meet7697
  - equipment_name: Confocal Microscope
    manufacturer_model: Howard-Thompson President7173
    settings_parameters: "7261 x g, 12\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Reynolds-Lynch Including2038
    settings_parameters: "10302 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      left.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      sense.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 303
    temperature_celsius: 32
    replicates: 2
  - step_description: Cells were transferred with ripa buffer to facilitate consumer.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 693
    temperature_celsius: 33
    replicates: 2
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Marc Dickerson
  and results were consistent across multiple biological replicates.
