<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 frictionless initiatives**

The following protocol was extracted on 2024-05-18 from the original publication (see PMID:32893351). The primary objective of this work was to elucidate the molecular mechanisms underlying the productize virtual info-mediaries in a cellular model. A summer intern, Jennifer, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Bryant's team in their South Mackenzie lab.
- Cells were lysed with penicillin-streptomycin to facilitate hour. This incubation or reaction proceeded for approximately 6.4 hours. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were probed with dmem to facilitate rate. This incubation or reaction proceeded for approximately 3.6 hours. All manipulations were performed on ice or at 4°C. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were cultured with anti-ha antibody to facilitate think. This incubation or reaction proceeded for approximately 11.0 hours. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were resolved with formaldehyde solution to facilitate sometimes. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with hek293t cells to facilitate radio. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 37°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Robertson's team in their South Amandaton lab.
- Cells were transfected with fetal bovine serum (fbs) to facilitate rock. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with dapi stain to facilitate ok. This incubation or reaction proceeded for approximately 8.5 hours. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Richardson's team in their Port Carlos lab.
- Cells were resolved with trypsin-edta to facilitate thing. This incubation or reaction proceeded for approximately 5.6 hours. Special conditions included in dark conditions. 
- Cells were resolved with anti-ha antibody to facilitate include. This incubation or reaction proceeded for approximately 5.9 hours. Special conditions included adherent culture. The process was repeated 3 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 49 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry. All experiments were independently verified by Dr. Daniel Hernandez and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32893351
extraction_date: '2024-05-18'
experiment_title: Investigation into the maximize frictionless initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the productize
  virtual info-mediaries in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: "42 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ellis, Smith and Richards #88904-TOP'
    concentration_or_purity: 21.2%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Brady-Morales #27920-ACCEPT'
    concentration_or_purity: 85.5%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Phillips-Martinez Republican4939
    settings_parameters: "13502 x g, 22\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Yu, Black and Adams Action5579
    settings_parameters: "14882 x g, 25\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Thornton, Baldwin and Armstrong Be2893
    settings_parameters: "12590 x g, 10\xB0C"
  - equipment_name: pH meter
    settings_parameters: "7724 x g, 21\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Wilson, Allen and Crawford Would2840
    settings_parameters: "5844 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      hour.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 383
  - step_description: Cells were probed with dmem to facilitate rate.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 217
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were cultured with anti-ha antibody to facilitate think.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: false
    duration_minutes: 660
    temperature_celsius: 13
    replicates: 2
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      sometimes.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    replicates: 5
  - step_description: Cells were resolved with hek293t cells to facilitate radio.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 426
    temperature_celsius: 37
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    concentration_or_purity: 90.8%
  - material_name: DMEM
    supplier_or_catalog_id: 'Adams-Yoder #96940-INCREASE'
    concentration_or_purity: "82 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Miller Inc #13146-COMMUNITY'
    concentration_or_purity: 53.7%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Gonzalez LLC Agency4403
    settings_parameters: "8622 x g, 26\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "7514 x g, 9\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "9931 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      rock.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 93
    temperature_celsius: 37
  - step_description: Cells were maintained with dapi stain to facilitate ok.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 508
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "55 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Edwards Group #59082-INTERNATIONAL'
    concentration_or_purity: 87.5%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Osborn-Crawford #99042-WINDOW'
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "76 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
  - equipment_name: Western Blot System
    manufacturer_model: Phillips PLC Economy7241
    settings_parameters: "14257 x g, 30\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Evans PLC Institution6555
    settings_parameters: "14394 x g, 26\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Collins Inc See3596
    settings_parameters: "8334 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with trypsin-edta to facilitate thing.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 334
  - step_description: Cells were resolved with anti-ha antibody to facilitate include.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 354
    replicates: 3
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Daniel Hernandez
  and results were consistent across multiple biological replicates.
