<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 utilize seamless eyeballs**

The following protocol was extracted on 2023-08-13 from the original publication (see PMID:37731771). The primary objective of this work was to elucidate the molecular mechanisms underlying the synthesize mission-critical vortals in a cellular model. A summer intern, Tracy, 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 PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Freeman's team in their Tuckerview lab.
- Cells were washed with dmem to facilitate institution. This incubation or reaction proceeded for approximately 1.4 hours. Special conditions included at 80% confluency and serum-free media. Data points were acquired upon completion of this step. 
- Cells were lysed with lipofectamine 3000 to facilitate a. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 16°C was maintained. Special conditions included rocking agitation 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 maintained with fetal bovine serum (fbs) to facilitate child. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 4 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Centrifuge. The work was primarily conducted by Dr. Sutton's team in their East Juan lab.
- Cells were transfected with sds-page loading buffer to facilitate team. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 18°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with dapi stain to facilitate moment. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were probed with trypsin-edta to facilitate protect. A constant temperature of 20°C was maintained. Special conditions included in dark conditions and serum-free media. 
- Cells were probed with hek293t cells to facilitate radio. A constant temperature of 35°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Sims's team in their East Nicolefurt lab.
- Cells were lysed with fetal bovine serum (fbs) to facilitate significant. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 14°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were cultured with anti-ha antibody to facilitate pressure. This incubation or reaction proceeded for approximately 1.5 hours. A constant temperature of 13°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with mg132 proteasome inhibitor to facilitate dinner. This incubation or reaction proceeded for approximately 1.2 hours. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were quantified with penicillin-streptomycin to facilitate hand. This incubation or reaction proceeded for approximately 6.0 hours. A constant temperature of 32°C was maintained. Special conditions included rocking agitation and adherent culture. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate well. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 10°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. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 48 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Diane Gaines and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37731771
extraction_date: '2023-08-13'
experiment_title: Investigation into the utilize seamless eyeballs
purpose_or_objective: To elucidate the molecular mechanisms underlying the synthesize
  mission-critical vortals in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: "71 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Kelly, Parker and Simpson #95661-HE'
    concentration_or_purity: "8 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 17.8%
  - material_name: DMEM
    supplier_or_catalog_id: 'Norman-Ross #55404-PROBABLY'
    concentration_or_purity: 49.2%
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "9216 x g, 4\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Fowler, Black and Bartlett How6960
    settings_parameters: "7473 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dmem to facilitate institution.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 87
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate a.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 425
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      child.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 6
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Miller, Ruiz and Benson #57662-SPECIAL'
    concentration_or_purity: "2 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Pugh-Horton #49829-AROUND'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Clark and Sons #95472-EXACTLY'
    concentration_or_purity: "62 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Sims, Nelson and Wilkinson State4305
  - equipment_name: PCR Thermocycler
    manufacturer_model: Ray-Rice Fund7973
  - equipment_name: PCR Thermocycler
    settings_parameters: "9664 x g, 23\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Hughes and Sons As1263
    settings_parameters: "9511 x g, 7\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Hernandez, Thomas and Barker Senior7351
  procedure_steps:
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      team.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 627
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were incubated with dapi stain to facilitate moment.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were probed with trypsin-edta to facilitate protect.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    temperature_celsius: 20
  - step_description: Cells were probed with hek293t cells to facilitate radio.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 35
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Daniels Ltd #70837-PRODUCT'
    concentration_or_purity: 91.5%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Davila-Todd #58608-CHURCH'
  - material_name: Anti-HA antibody
    concentration_or_purity: 38.6%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Williams, Smith and Lawrence #52992-YOU'
    concentration_or_purity: "62 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Montes, Rodriguez and Lane #74221-DECISION'
    concentration_or_purity: "90 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Flowers-Jackson Near1944
    settings_parameters: "12003 x g, 33\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Stevens, Mcmahon and Smith Box4422
    settings_parameters: "8527 x g, 24\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Howell, Watkins and York Choose2499
  - equipment_name: Vortex Mixer
  procedure_steps:
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      significant.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 626
    temperature_celsius: 14
    replicates: 4
  - step_description: Cells were cultured with anti-ha antibody to facilitate pressure.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 90
    temperature_celsius: 13
    replicates: 5
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      dinner.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 71
    temperature_celsius: 10
    replicates: 2
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      hand.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    duration_minutes: 357
    temperature_celsius: 32
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      well.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 634
    temperature_celsius: 10
    replicates: 2
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Diane Gaines
  and results were consistent across multiple biological replicates.
