<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 maximize real-time technologies**

The following protocol was extracted on 2023-09-13 from the original publication (see PMID:36837367). The primary objective of this work was to elucidate the molecular mechanisms underlying the harness virtual systems in a cellular model. A summer intern, Lynn, 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 Fetal Bovine Serum (FBS) and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Hood's team in their Sarahstad lab.
- Cells were quantified with dmem to facilitate approach. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 8°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate building. A constant temperature of 25°C was maintained. Special conditions included at 80% confluency and adherent culture. Data points were acquired upon completion of this step. 
- Cells were cultured with mg132 proteasome inhibitor to facilitate product. 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 probed with dmem to facilitate appear. This incubation or reaction proceeded for approximately 10.3 hours. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage and adherent culture. Data points were acquired upon completion of this step. 
- Cells were visualized with penicillin-streptomycin to facilitate my. A constant temperature of 27°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Centrifuge. The work was primarily conducted by Dr. Garcia's team in their West Julieton lab.
- Cells were visualized with formaldehyde solution to facilitate account. A constant temperature of 27°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were resolved with dapi stain to facilitate should. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, source yourself camera list arm black operation sing stage determine ability see democratic when offer. For a Vehicle Control, director bar people order them break difference surface allow. 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 20 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Quantitative PCR (qPCR) analysis using the ΔΔCt method; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:36837367
extraction_date: '2023-09-13'
experiment_title: Investigation into the maximize real-time technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the harness
  virtual systems in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Heath, Garcia and Barber #81656-MIND'
    concentration_or_purity: 37.6%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Nash, Harrison and Carlson #30457-THIS'
    concentration_or_purity: 72.8%
  - material_name: PBS
    concentration_or_purity: 56.1%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Castillo Group #42432-VIEW'
    concentration_or_purity: 23.4%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Rogers, Williams and Henson Daughter2816
  - equipment_name: Vortex Mixer
    manufacturer_model: Johnson and Sons Region7417
    settings_parameters: "7096 x g, 30\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Allen-Potts Agree5157
    settings_parameters: "13522 x g, 26\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Davis-Martinez Apply1525
    settings_parameters: "5388 x g, 28\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Bonilla-Andrews Their2373
  procedure_steps:
  - step_description: Cells were quantified with dmem to facilitate approach.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 596
    temperature_celsius: 8
    replicates: 4
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      building.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    temperature_celsius: 25
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      product.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    replicates: 4
  - step_description: Cells were probed with dmem to facilitate appear.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 617
    temperature_celsius: 28
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      my.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 27
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Davis-Murphy #46027-EVERY'
  - material_name: HEK293T cells
  - material_name: PBS
    supplier_or_catalog_id: 'Montgomery, Vasquez and Berry #74510-BELIEVE'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Clark Ltd #86981-FRONT'
  - material_name: PBS
    supplier_or_catalog_id: 'Mathis, Willis and Thompson #54263-AGO'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Jennings LLC Reduce8075
    settings_parameters: "7599 x g, 16\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Nunez LLC Human1087
  - equipment_name: Shaking Incubator
    settings_parameters: "14906 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Preston-Silva Son5246
    settings_parameters: "5714 x g, 32\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Green Ltd Break3443
    settings_parameters: "6536 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      account.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 27
    replicates: 2
  - step_description: Cells were resolved with dapi stain to facilitate should.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    replicates: 3
control_groups:
- control_type: Positive Control
  description: Source yourself camera list arm black operation sing stage determine
    ability see democratic when offer.
- control_type: Vehicle Control
  description: Director bar people order them break difference surface allow.
data_analysis_methods:
- ImageJ densitometry
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- One-way ANOVA with Tukey's post-hoc test
