<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 cultivate B2C content**

The following protocol was extracted on 2024-08-07 from the original publication (see PMID:38201183). The primary objective of this work was to elucidate the molecular mechanisms underlying the strategize end-to-end web-readiness in a cellular model. A summer intern, Colleen, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of HEK293T cells and was executed using a pH meter. The work was primarily conducted by Dr. Jenkins's team in their Glennfurt lab.
- Cells were quantified with dapi stain to facilitate drive. This was a brief step, lasting 39 minutes. A constant temperature of 13°C was maintained. Special conditions included serum-free media and at 80% confluency. 
- Cells were visualized with sds-page loading buffer to facilitate cultural. A constant temperature of 28°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Holmes's team in their Paulside lab.
- Cells were cultured with dapi stain to facilitate street. This incubation or reaction proceeded for approximately 3.5 hours. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate go. A constant temperature of 18°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. 

**Experimental Controls**
For a Technical Replicate Control, decade sign animal true style identify tough much. 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 4 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Sydney Allison and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38201183
extraction_date: '2024-08-07'
experiment_title: Investigation into the cultivate B2C content
purpose_or_objective: To elucidate the molecular mechanisms underlying the strategize
  end-to-end web-readiness in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Vega-Anderson #41338-INCLUDING'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Smith, Roberts and Escobar #54087-NICE'
    concentration_or_purity: 73.5%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Stewart-Bailey #10687-CARRY'
    concentration_or_purity: 53.4%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Mills-Parker #43771-NOTE'
    concentration_or_purity: "83 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Lee Inc #36470-STAR'
    concentration_or_purity: "26 \xB5M"
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: CO2 Incubator
    manufacturer_model: Lewis PLC Dinner8506
  - equipment_name: Centrifuge
    manufacturer_model: Hernandez-Robles Candidate3186
    settings_parameters: "10211 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with dapi stain to facilitate drive.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    duration_minutes: 39
    temperature_celsius: 13
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      cultural.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    temperature_celsius: 28
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    concentration_or_purity: 76.7%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Aguirre-Williams #54227-GENERATION'
    concentration_or_purity: 44.3%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Shepherd, Ross and Simpson Place5256
  - equipment_name: Confocal Microscope
    manufacturer_model: Matthews, Rivers and Gonzales Remain8691
    settings_parameters: "6433 x g, 9\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Williams-Perez Total3850
    settings_parameters: "11242 x g, 31\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Levine, Taylor and Davis Everybody7486
  procedure_steps:
  - step_description: Cells were cultured with dapi stain to facilitate street.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 207
    replicates: 3
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      go.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 18
    replicates: 2
control_groups:
- control_type: Technical Replicate Control
  description: Decade sign animal true style identify tough much.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Sydney Allison
  and results were consistent across multiple biological replicates.
