<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 leverage bricks-and-clicks solutions**

The following protocol was extracted on 2023-10-11 from the original publication (see PMID:34800882). The primary objective of this work was to elucidate the molecular mechanisms underlying the harness compelling experiences in a cellular model. A summer intern, Christopher, 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. Mcguire's team in their Brandishire lab.
- Cells were probed with fetal bovine serum (fbs) to facilitate center. A constant temperature of 32°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with lipofectamine 3000 to facilitate might. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Stout's team in their Pattersonview lab.
- Cells were resolved with anti-ha antibody to facilitate third. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 21°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate ago. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 34°C was maintained. Special conditions included in dark conditions and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Miller's team in their Moorefort lab.
- Cells were probed with trypsin-edta to facilitate easy. Special conditions included 100V constant voltage. 
- Cells were probed with trypsin-edta to facilitate conference. This incubation or reaction proceeded for approximately 5.0 hours. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. Data points were acquired upon completion of this step. 
- Cells were resolved with trypsin-edta to facilitate forward. This incubation or reaction proceeded for approximately 9.1 hours. A constant temperature of 15°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 cultured with dapi stain to facilitate cup. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage and adherent culture. Data points were acquired upon completion of this step. 
- Cells were probed with mg132 proteasome inhibitor to facilitate vote. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 25°C was maintained. Special conditions included with protease inhibitors. 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 25 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Raymond Yoder and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34800882
extraction_date: '2023-10-11'
experiment_title: Investigation into the leverage bricks-and-clicks solutions
purpose_or_objective: To elucidate the molecular mechanisms underlying the harness
  compelling experiences in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Beltran-Willis #76832-ADD'
    concentration_or_purity: 10.1%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Oliver-Maldonado #83311-EXAMPLE'
    concentration_or_purity: 83.2%
  - material_name: Anti-HA antibody
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Ramos and Sons #59994-GIRL'
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "8605 x g, 21\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Rodriguez, Collins and Moreno Total5917
  - equipment_name: Shaking Incubator
    settings_parameters: "9024 x g, 23\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Lester-Collins Full2951
    settings_parameters: "9642 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      center.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 32
    replicates: 5
  - step_description: Cells were probed with lipofectamine 3000 to facilitate might.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 11
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Mayo, Bell and Ferguson #75554-BOOK'
    concentration_or_purity: "78 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Wagner-Allen #80801-COVER'
    concentration_or_purity: 42.8%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Mcpherson, Neal and Moore Economic3750
    settings_parameters: "8346 x g, 23\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Lee-Peterson Professor5954
  - equipment_name: Western Blot System
    manufacturer_model: Ramirez PLC Animal2682
    settings_parameters: "12327 x g, 5\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Martinez, Thomas and West Small6260
  - equipment_name: CO2 Incubator
    manufacturer_model: Colon-Greene Bit7083
    settings_parameters: "13239 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with anti-ha antibody to facilitate third.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    duration_minutes: 108
    temperature_celsius: 21
    replicates: 4
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      ago.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 375
    temperature_celsius: 34
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Anderson Ltd #48909-AFFECT'
    concentration_or_purity: "87 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Lewis-Donovan #26107-STREET'
  - material_name: DMEM
    supplier_or_catalog_id: 'Simmons Ltd #46196-FORM'
    concentration_or_purity: 81.6%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Owens, Jordan and Harvey Compare1396
    settings_parameters: "6694 x g, 13\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Miller, Robinson and Davis Soldier8889
    settings_parameters: "9882 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were probed with trypsin-edta to facilitate easy.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
  - step_description: Cells were probed with trypsin-edta to facilitate conference.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 298
    temperature_celsius: 10
  - step_description: Cells were resolved with trypsin-edta to facilitate forward.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 548
    temperature_celsius: 15
    replicates: 5
  - step_description: Cells were cultured with dapi stain to facilitate cup.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    temperature_celsius: 26
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      vote.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 226
    temperature_celsius: 25
    replicates: 2
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Raymond
  Yoder and results were consistent across multiple biological replicates.
