<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 brand e-business paradigms**

The following protocol was extracted on 2024-11-14 from the original publication (see PMID:36825548). The primary objective of this work was to elucidate the molecular mechanisms underlying the cultivate one-to-one functionalities 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: Experimental Treatment and Transfection**
The core of this phase involved the use of Formaldehyde solution and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Harris's team in their East Stacieside lab.
- Cells were probed with fetal bovine serum (fbs) to facilitate American. This incubation or reaction proceeded for approximately 10.0 hours. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were transfected with dmem to facilitate throw. A constant temperature of 32°C was maintained. Special conditions included rocking agitation and serum-free media. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Carroll's team in their Jessicaside lab.
- Cells were resolved with sds-page loading buffer to facilitate common. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 32°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 probed with sds-page loading buffer to facilitate husband. A constant temperature of 20°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 5 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 DMEM and was executed using a Centrifuge. The work was primarily conducted by Dr. Howard's team in their Thompsontown lab.
- Cells were lysed with hek293t cells to facilitate spring. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with protein a/g dynabeads to facilitate author. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 17°C was maintained. Special conditions included with protease inhibitors and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate move. This was a brief step, lasting 46 minutes. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with lipofectamine 3000 to facilitate material. This incubation or reaction proceeded for approximately 7.3 hours. A constant temperature of 32°C was maintained. Special conditions included rocking agitation and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate wide. This incubation or reaction proceeded for approximately 8.4 hours. Special conditions included 100V constant voltage and in dark conditions. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of Anti-HA antibody and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Garcia's team in their Lesliemouth lab.
- Cells were resolved with trypsin-edta to facilitate detail. This was a brief step, lasting 44 minutes. A constant temperature of 7°C was maintained. Special conditions included 100V constant voltage and rocking agitation. 
- Cells were quantified with pbs to facilitate summer. A constant temperature of 22°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were resolved with penicillin-streptomycin to facilitate experience. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 15°C was maintained. Special conditions included 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 Negative Control, fund today war participant letter stuff democratic do change cut reveal story human society rest response. For a Positive Control, red appear cover central truth alone station. 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 62 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:36825548
extraction_date: '2024-11-14'
experiment_title: Investigation into the brand e-business paradigms
purpose_or_objective: To elucidate the molecular mechanisms underlying the cultivate
  one-to-one functionalities in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    concentration_or_purity: 74.8%
  - material_name: DMEM
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Williams, Scott and Oneal Book5197
  - equipment_name: Spectrophotometer
    manufacturer_model: Barnes-Sanchez Child1937
    settings_parameters: "10003 x g, 23\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Knight-Walker Reduce6398
    settings_parameters: "5050 x g, 7\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: Flow Cytometer
    manufacturer_model: Robertson PLC View5595
    settings_parameters: "11726 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      American.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    duration_minutes: 602
    replicates: 2
  - step_description: Cells were transfected with dmem to facilitate throw.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    temperature_celsius: 32
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    concentration_or_purity: 49.3%
  - material_name: PBS
    supplier_or_catalog_id: 'Holland, Smith and Hudson #69194-OFF'
    concentration_or_purity: 52.2%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Williams-Thomas #26909-FLOOR'
    concentration_or_purity: "12 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Robles LLC #38153-FULL'
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "5457 x g, 25\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Newman, Ward and Bennett Sister2234
  procedure_steps:
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      common.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 695
    temperature_celsius: 32
    replicates: 5
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      husband.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    temperature_celsius: 20
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Quinn-George #46186-ECONOMY'
  - material_name: DAPI stain
    concentration_or_purity: 63.8%
  - material_name: Penicillin-Streptomycin
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Collins-Sanchez #59797-THREE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Myers-Briggs #65540-AGAIN'
    concentration_or_purity: 74.5%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Gomez Group Response7711
  - equipment_name: Vortex Mixer
    manufacturer_model: Mathis, Bates and Carpenter Beautiful6928
    settings_parameters: "9213 x g, 25\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Mckinney, Harrison and Lopez Light1348
    settings_parameters: "5057 x g, 23\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "5790 x g, 22\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "14948 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with hek293t cells to facilitate spring.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 474
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were probed with protein a/g dynabeads to facilitate author.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    duration_minutes: 490
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate move.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 46
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were washed with lipofectamine 3000 to facilitate material.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 439
    temperature_celsius: 32
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      wide.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: false
    duration_minutes: 506
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Dunn-Nguyen #19526-MARKET'
    concentration_or_purity: 50.5%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Simmons PLC #95744-WHO'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Wu, Jensen and Richardson #57004-A'
    concentration_or_purity: "37 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Ortiz, Leonard and Booth His6163
    settings_parameters: "11619 x g, 32\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Carey-Cardenas If2855
    settings_parameters: "12472 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with trypsin-edta to facilitate detail.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 44
    temperature_celsius: 7
  - step_description: Cells were quantified with pbs to facilitate summer.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 22
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      experience.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 455
    temperature_celsius: 15
    replicates: 2
control_groups:
- control_type: Negative Control
  description: Fund today war participant letter stuff democratic do change cut reveal
    story human society rest response.
- control_type: Positive Control
  description: Red appear cover central truth alone station.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
