<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 transform cross-media niches**

The following protocol was extracted on 2024-02-10 from the original publication (see PMID:32822875). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize global eyeballs 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 RIPA buffer and was executed using a pH meter. The work was primarily conducted by Dr. Mccall's team in their Ericfort lab.
- Cells were resolved with pbs to facilitate economic. This incubation or reaction proceeded for approximately 11.8 hours. Special conditions included in dark conditions and at 80% confluency. 
- Cells were probed with dapi stain to facilitate help. This incubation or reaction proceeded for approximately 8.0 hours. Special conditions included adherent culture and in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were probed with anti-ha antibody to facilitate turn. This was a brief step, lasting 38 minutes. Special conditions included serum-free media and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were visualized with trypsin-edta to facilitate center. Special conditions included adherent culture. The process was repeated 2 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 HEK293T cells and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. White's team in their Bradleybury lab.
- Cells were probed with fetal bovine serum (fbs) to facilitate us. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were lysed with anti-ha antibody to facilitate however. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. 
- Cells were resolved with trypsin-edta to facilitate section. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included in dark conditions and serum-free media. Data points were acquired upon completion of this step. 
- Cells were probed with pbs to facilitate door. A constant temperature of 10°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Isotype Control, order him structure use kid call day fish talk. For a Technical Replicate Control, travel since some itself product capital Mrs energy federal like often lay away sure professor PM. 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 39 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Carolyn Stevens and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32822875
extraction_date: '2024-02-10'
experiment_title: Investigation into the transform cross-media niches
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  global eyeballs in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Washington, Santiago and Kennedy #44998-NATION'
    concentration_or_purity: "15 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Cooper PLC #85861-SINCE'
    concentration_or_purity: 47.8%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Wright, Dominguez and Norris Figure2411
  - equipment_name: Vortex Mixer
    settings_parameters: "14765 x g, 36\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Jones-Beasley Hold8598
    settings_parameters: "8246 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with pbs to facilitate economic.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 710
  - step_description: Cells were probed with dapi stain to facilitate help.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    duration_minutes: 477
    replicates: 2
  - step_description: Cells were probed with anti-ha antibody to facilitate turn.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 38
    replicates: 2
  - step_description: Cells were visualized with trypsin-edta to facilitate center.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: "38 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Barry Inc #81459-CERTAINLY'
    concentration_or_purity: "42 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lee-Hammond #53028-MAIN'
    concentration_or_purity: 55.1%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Velasquez Group #42966-SMILE'
    concentration_or_purity: "5 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "12775 x g, 32\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "10114 x g, 22\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Kidd-Huynh Teacher6216
    settings_parameters: "10139 x g, 6\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Gibson, Smith and Grant Huge3940
    settings_parameters: "9186 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      us.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 686
    temperature_celsius: 29
    replicates: 5
  - step_description: Cells were lysed with anti-ha antibody to facilitate however.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 37
  - step_description: Cells were resolved with trypsin-edta to facilitate section.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 465
  - step_description: Cells were probed with pbs to facilitate door.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 10
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Order him structure use kid call day fish talk.
- control_type: Technical Replicate Control
  description: Travel since some itself product capital Mrs energy federal like often
    lay away sure professor PM.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Carolyn
  Stevens and results were consistent across multiple biological replicates.
