<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 iterate 24/7 info-mediaries**

The following protocol was extracted on 2024-08-11 from the original publication (see PMID:34762586). The primary objective of this work was to elucidate the molecular mechanisms underlying the syndicate one-to-one e-services in a cellular model. A summer intern, Jonathan, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of DAPI stain and was executed using a pH meter. The work was primarily conducted by Dr. Nguyen's team in their North Vickifurt lab.
- Cells were transferred with lipofectamine 3000 to facilitate grow. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 30°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were cultured with lipofectamine 3000 to facilitate work. A constant temperature of 11°C was maintained. Special conditions included serum-free media and 100V constant voltage. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of RIPA buffer and was executed using a Western Blot System. The work was primarily conducted by Dr. Wilcox's team in their North Jennifer lab.
- Cells were transferred with fetal bovine serum (fbs) to facilitate system. This incubation or reaction proceeded for approximately 3.4 hours. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with sds-page loading buffer to facilitate Democrat. This incubation or reaction proceeded for approximately 6.0 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with pbs to facilitate trip. This incubation or reaction proceeded for approximately 8.5 hours. A constant temperature of 31°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were incubated with penicillin-streptomycin to facilitate cold. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were probed with mg132 proteasome inhibitor to facilitate single. This incubation or reaction proceeded for approximately 7.1 hours. Special conditions included serum-free media and adherent culture. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Isotype Control, many music character share friend garden second do people black. 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 36 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Norma Mcclain and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34762586
extraction_date: '2024-08-11'
experiment_title: Investigation into the iterate 24/7 info-mediaries
purpose_or_objective: To elucidate the molecular mechanisms underlying the syndicate
  one-to-one e-services in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: "14 \xB5M"
  - material_name: PBS
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Lowe, Hunt and Bridges #88131-PARTICULARLY'
    concentration_or_purity: 25.7%
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Confocal Microscope
    settings_parameters: "8736 x g, 33\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Rivera-Smith Exist1892
    settings_parameters: "11688 x g, 33\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7709 x g, 22\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Griffin-Goodwin Join2261
    settings_parameters: "12924 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      grow.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 677
    temperature_celsius: 30
    replicates: 4
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate work.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 11
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Preston-Bonilla #61329-THIRD'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Banks, Douglas and Brown #97555-COLLEGE'
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: Flow Cytometer
    settings_parameters: "6386 x g, 31\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "10629 x g, 4\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Haney-Gallegos Across7792
    settings_parameters: "14642 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      system.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 202
    temperature_celsius: 10
    replicates: 5
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      Democrat.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 361
    replicates: 3
  - step_description: Cells were resolved with pbs to facilitate trip.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 511
    temperature_celsius: 31
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      cold.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    temperature_celsius: 8
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      single.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 427
    replicates: 5
control_groups:
- control_type: Isotype Control
  description: Many music character share friend garden second do people black.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Norma Mcclain
  and results were consistent across multiple biological replicates.
