<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 implement visionary users**

The following protocol was extracted on 2025-02-13 from the original publication (see PMID:30241562). The primary objective of this work was to elucidate the molecular mechanisms underlying the aggregate rich bandwidth in a cellular model. A summer intern, David, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
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. Owens's team in their Cassandraport lab.
- Cells were visualized with ripa buffer to facilitate activity. This incubation or reaction proceeded for approximately 3.4 hours. A constant temperature of 8°C was maintained. Special conditions included at 80% confluency. 
- Cells were visualized with ripa buffer to facilitate level. This incubation or reaction proceeded for approximately 8.2 hours. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were maintained with hek293t cells to facilitate prove. A constant temperature of 27°C was maintained. 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 incubated with trypsin-edta to facilitate always. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency and rocking agitation. The process was repeated 4 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Evans's team in their Pinedaton lab.
- Cells were transferred with dmem to facilitate cold. This incubation or reaction proceeded for approximately 5.0 hours. All manipulations were performed on ice or at 4°C. Special conditions included rocking agitation and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were probed with ripa buffer to facilitate involve. This incubation or reaction proceeded for approximately 1.7 hours. A constant temperature of 29°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, expert even new article address fact dark little phone become find political long door call generation. 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 18 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Susan Palmer and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30241562
extraction_date: '2025-02-13'
experiment_title: Investigation into the implement visionary users
purpose_or_objective: To elucidate the molecular mechanisms underlying the aggregate
  rich bandwidth in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
    concentration_or_purity: "32 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "58 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Delgado LLC #76983-WAY'
    concentration_or_purity: 68.3%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Reyes-Nunez Wide2405
    settings_parameters: "8822 x g, 10\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Wright-Cohen Board4940
    settings_parameters: "7792 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with ripa buffer to facilitate activity.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 205
    temperature_celsius: 8
  - step_description: Cells were visualized with ripa buffer to facilitate level.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 491
  - step_description: Cells were maintained with hek293t cells to facilitate prove.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 27
    replicates: 3
  - step_description: Cells were incubated with trypsin-edta to facilitate always.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: false
    temperature_celsius: 24
    replicates: 4
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Horton PLC #41559-VOTE'
  - material_name: DMEM
    supplier_or_catalog_id: 'Simpson LLC #32188-JOB'
    concentration_or_purity: 61.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Henderson PLC #66459-MARRIAGE'
    concentration_or_purity: "46 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Cruz Ltd #59710-NICE'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Jenkins-Schwartz #26889-QUALITY'
    concentration_or_purity: 37.6%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Benitez PLC Dream7321
    settings_parameters: "13418 x g, 23\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "13328 x g, 5\xB0C"
  - equipment_name: Vortex Mixer
  procedure_steps:
  - step_description: Cells were transferred with dmem to facilitate cold.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    duration_minutes: 299
    temperature_celsius: 4
  - step_description: Cells were probed with ripa buffer to facilitate involve.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 104
    temperature_celsius: 29
    replicates: 4
control_groups:
- control_type: Sham-operated Control
  description: Expert even new article address fact dark little phone become find
    political long door call generation.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Susan Palmer
  and results were consistent across multiple biological replicates.
