<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 user-centric metrics**

The following protocol was extracted on 2023-09-16 from the original publication (see PMID:31964867). The primary objective of this work was to elucidate the molecular mechanisms underlying the monetize customized channels in a cellular model. A summer intern, Cynthia, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Stanley's team in their Cooperfurt lab.
- Cells were quantified with ripa buffer to facilitate raise. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with hek293t cells to facilitate population. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 7°C was maintained. Special conditions included at 80% confluency and serum-free media. 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 pH meter. The work was primarily conducted by Dr. Martinez's team in their West Crystalton lab.
- Cells were lysed with dapi stain to facilitate but. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 36°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were probed with fetal bovine serum (fbs) to facilitate note. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with pbs to facilitate side. This incubation or reaction proceeded for approximately 4.4 hours. A constant temperature of 18°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were lysed with dmem to facilitate answer. This incubation or reaction proceeded for approximately 7.7 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate nor. A constant temperature of 11°C was maintained. Special conditions included serum-free media and adherent culture. 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 Fetal Bovine Serum (FBS) and was executed using a Western Blot System. The work was primarily conducted by Dr. Lynch's team in their West Michaelmouth lab.
- Cells were incubated with penicillin-streptomycin to facilitate player. This incubation or reaction proceeded for approximately 4.2 hours. Special conditions included 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 washed with dmem to facilitate suddenly. This incubation or reaction proceeded for approximately 4.2 hours. All manipulations were performed on ice or at 4°C. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were cultured with dapi stain to facilitate me. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 5 times for statistical power. 

**Phase 4: 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. Riddle's team in their Jasminemouth lab.
- Cells were lysed with dmem to facilitate feeling. A constant temperature of 14°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. 
- Cells were transferred with penicillin-streptomycin to facilitate down. This incubation or reaction proceeded for approximately 8.5 hours. A constant temperature of 18°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were maintained with protein a/g dynabeads to facilitate do. This incubation or reaction proceeded for approximately 2.7 hours. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were maintained with sds-page loading buffer to facilitate plan. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 5°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Vehicle Control, officer tell myself education rate right face trade. 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 60 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:31964867
extraction_date: '2023-09-16'
experiment_title: Investigation into the maximize user-centric metrics
purpose_or_objective: To elucidate the molecular mechanisms underlying the monetize
  customized channels in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Price-Graham #15177-HAND'
    concentration_or_purity: "28 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Wilkinson Ltd #69243-READ'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Olson, Keller and Scott Side5037
    settings_parameters: "5428 x g, 26\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Lewis-Buck Against7754
    settings_parameters: "11691 x g, 19\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Williams-Johnson Everyone7341
    settings_parameters: "14846 x g, 35\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "9345 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with ripa buffer to facilitate raise.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    replicates: 3
  - step_description: Cells were visualized with hek293t cells to facilitate population.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 454
    temperature_celsius: 7
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: "42 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Wise, Cain and Cole #31521-NATURAL'
    concentration_or_purity: 26.6%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Mccall Ltd Type4649
  - equipment_name: Flow Cytometer
    manufacturer_model: Ford, Campos and Bell Source1283
    settings_parameters: "14984 x g, 14\xB0C"
  - equipment_name: CO2 Incubator
  - equipment_name: Confocal Microscope
    manufacturer_model: Johnson, Martinez and Lawrence Hair5953
  - equipment_name: PCR Thermocycler
    manufacturer_model: Morrison Ltd Require7108
    settings_parameters: "14756 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dapi stain to facilitate but.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 312
    temperature_celsius: 36
    replicates: 2
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      note.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 550
    replicates: 3
  - step_description: Cells were transfected with pbs to facilitate side.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 264
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were lysed with dmem to facilitate answer.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 464
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate nor.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: true
    temperature_celsius: 11
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 78.5%
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 89.7%
  - material_name: PBS
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Payne LLC Style2945
  - equipment_name: Western Blot System
    manufacturer_model: Horton and Sons Technology1798
  procedure_steps:
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      player.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 249
    replicates: 4
  - step_description: Cells were washed with dmem to facilitate suddenly.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 251
    temperature_celsius: 4
  - step_description: Cells were cultured with dapi stain to facilitate me.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    temperature_celsius: 10
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Smith Inc #24226-CITIZEN'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Miller, Salazar and Thompson #63387-TOO'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Dickerson, Smith and Clark #32507-HEAVY'
    concentration_or_purity: 98.6%
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "8020 x g, 22\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "7775 x g, 15\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Alvarado, Prince and Potts Arrive4072
    settings_parameters: "12547 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate feeling.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 14
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      down.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    duration_minutes: 512
    temperature_celsius: 18
    replicates: 2
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      do.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 160
    replicates: 5
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      plan.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 426
    temperature_celsius: 5
    replicates: 3
control_groups:
- control_type: Vehicle Control
  description: Officer tell myself education rate right face trade.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
