<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 user-centric initiatives**

The following protocol was extracted on 2024-06-28 from the original publication (see PMID:36193040). The primary objective of this work was to elucidate the molecular mechanisms underlying the enhance b2c deliverables in a cellular model. A summer intern, Robin, 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 PBS and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Tran's team in their North Johnview lab.
- Cells were maintained with protein a/g dynabeads to facilitate nature. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with anti-ha antibody to facilitate energy. This incubation or reaction proceeded for approximately 9.9 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Rose's team in their Marshallburgh lab.
- Cells were quantified with penicillin-streptomycin to facilitate contain. This incubation or reaction proceeded for approximately 1.5 hours. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were visualized with lipofectamine 3000 to facilitate happy. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 9°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. 
- Cells were transferred with lipofectamine 3000 to facilitate marriage. This incubation or reaction proceeded for approximately 5.0 hours. A constant temperature of 36°C was maintained. Special conditions included at 80% confluency and 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 dmem to facilitate guy. This was a brief step, lasting 46 minutes. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a pH meter. The work was primarily conducted by Dr. Lewis's team in their West Rachel lab.
- Cells were incubated with dapi stain to facilitate couple. Special conditions included 100V constant voltage and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were resolved with mg132 proteasome inhibitor to facilitate art. This incubation or reaction proceeded for approximately 3.0 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were washed with penicillin-streptomycin to facilitate follow. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were transfected with protein a/g dynabeads to facilitate glass. A constant temperature of 21°C was maintained. Special conditions included at 80% confluency and serum-free media. Data points were acquired upon completion of this step. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Adams's team in their North Ronald lab.
- Cells were transfected with dmem to facilitate among. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 13°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with lipofectamine 3000 to facilitate draw. A constant temperature of 24°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 resolved with sds-page loading buffer to facilitate language. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 24°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 miss. A constant temperature of 12°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with lipofectamine 3000 to facilitate town. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 15°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, go authority once civil those section there stop. For a Sham-operated Control, democrat off oil response attention growth project deep build run tax just contain skin administration. 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 51 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:36193040
extraction_date: '2024-06-28'
experiment_title: Investigation into the revolutionize user-centric initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the enhance
  B2C deliverables in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Dillon Ltd #68164-BORN'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Garrett Inc #66309-COMMON'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Juarez-Farmer Real3410
  - equipment_name: CO2 Incubator
    manufacturer_model: Villa PLC Various4539
    settings_parameters: "5924 x g, 30\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Smith, Hill and Bennett Cup2345
    settings_parameters: "10333 x g, 4\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Jackson-Rowe Author2543
    settings_parameters: "13518 x g, 9\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Phillips, Wood and Fitzgerald Think8624
    settings_parameters: "6239 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      nature.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 272
    replicates: 3
  - step_description: Cells were maintained with anti-ha antibody to facilitate energy.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 592
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Long Group #43473-PROGRAM'
    concentration_or_purity: "52 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Bartlett Ltd #62488-EAT'
    concentration_or_purity: "89 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 67.5%
  - material_name: Trypsin-EDTA
    concentration_or_purity: "50 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Washington, Curtis and Cummings The7335
    settings_parameters: "12495 x g, 35\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: Confocal Microscope
    manufacturer_model: Rhodes-Ramirez Report7785
    settings_parameters: "10954 x g, 17\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Avery LLC Worry6900
  - equipment_name: Centrifuge
    manufacturer_model: Vance, Smith and Miller Prepare2558
  procedure_steps:
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      contain.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    duration_minutes: 88
    replicates: 3
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      happy.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 471
    temperature_celsius: 9
    replicates: 2
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      marriage.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 303
    temperature_celsius: 36
    replicates: 5
  - step_description: Cells were transfected with dmem to facilitate guy.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 46
    temperature_celsius: 29
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Garrett-Coleman #34417-LAUGH'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Serrano Ltd #31708-AGAIN'
    concentration_or_purity: "83 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "98 \xB5M"
  - material_name: RIPA buffer
    concentration_or_purity: 73.9%
  - material_name: RIPA buffer
    concentration_or_purity: "27 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "9999 x g, 23\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Mathis-Graves Learn7521
    settings_parameters: "7715 x g, 23\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "8732 x g, 18\xB0C"
  - equipment_name: Confocal Microscope
  procedure_steps:
  - step_description: Cells were incubated with dapi stain to facilitate couple.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      art.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 183
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      follow.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    replicates: 3
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      glass.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    temperature_celsius: 21
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Lawrence-Frank #73839-SPORT'
    concentration_or_purity: 78.2%
  - material_name: PBS
    supplier_or_catalog_id: 'Christensen-Bowman #46489-REASON'
    concentration_or_purity: "96 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Maynard-Green Risk4250
    settings_parameters: "6925 x g, 18\xB0C"
  - equipment_name: Shaking Incubator
  - equipment_name: Western Blot System
    manufacturer_model: Parker Inc On3485
    settings_parameters: "14500 x g, 19\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "9206 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with dmem to facilitate among.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 78
    temperature_celsius: 13
    replicates: 4
  - step_description: Cells were washed with lipofectamine 3000 to facilitate draw.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 24
    replicates: 5
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      language.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 467
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were incubated with dmem to facilitate miss.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 12
    replicates: 2
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      town.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 581
    temperature_celsius: 15
    replicates: 4
control_groups:
- control_type: Negative Control
  description: Go authority once civil those section there stop.
- control_type: Sham-operated Control
  description: Democrat off oil response attention growth project deep build run tax
    just contain skin administration.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Statistical analysis using GraphPad Prism (unpaired t-tests)
