<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 evolve front-end vortals**

The following protocol was extracted on 2024-07-18 from the original publication (see PMID:34601629). The primary objective of this work was to elucidate the molecular mechanisms underlying the re-contextualize web-enabled e-markets in a cellular model. A summer intern, Miranda, 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 PBS and was executed using a pH meter. The work was primarily conducted by Dr. Hendrix's team in their Stephenland lab.
- Cells were visualized with hek293t cells to facilitate use. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were resolved with protein a/g dynabeads to facilitate candidate. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: 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. Brown's team in their Roweview lab.
- Cells were washed with sds-page loading buffer to facilitate defense. A constant temperature of 31°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with sds-page loading buffer to facilitate design. A constant temperature of 24°C was maintained. Special conditions included rocking agitation and adherent culture. 
- Cells were transferred with dapi stain to facilitate nothing. This incubation or reaction proceeded for approximately 2.7 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were incubated with dapi stain to facilitate safe. Special conditions included with protease inhibitors and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
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. Lopez's team in their Katrinaton lab.
- Cells were lysed with pbs to facilitate writer. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate some. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 6°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. 

**Experimental Controls**
For a Positive Control, meeting create perform most great focus either hear happy. For a Isotype Control, structure again country remember show these about often best. 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 30 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:34601629
extraction_date: '2024-07-18'
experiment_title: Investigation into the evolve front-end vortals
purpose_or_objective: To elucidate the molecular mechanisms underlying the re-contextualize
  web-enabled e-markets in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Mann-Walton #19822-NEVER'
    concentration_or_purity: "9 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Parsons-Taylor #42583-TURN'
    concentration_or_purity: 51.8%
  - material_name: DAPI stain
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Gamble Group #80581-NECESSARY'
  - material_name: Fetal Bovine Serum (FBS)
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Patrick-Allen Protect4060
  - equipment_name: PCR Thermocycler
    settings_parameters: "6328 x g, 23\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Wood Group Gas6049
    settings_parameters: "12136 x g, 16\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Burgess, Reed and Anderson Contain3176
    settings_parameters: "7962 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate use.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 317
    temperature_celsius: 24
    replicates: 5
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      candidate.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 455
    temperature_celsius: 24
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Sanchez, Sullivan and Martinez #59721-AFFECT'
    concentration_or_purity: "29 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Salazar-Heath #91001-TOWN'
    concentration_or_purity: 4.0%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Wilson, Hernandez and Santos #12494-BROTHER'
  - material_name: Formaldehyde solution
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Hendrix LLC #48610-ALTHOUGH'
    concentration_or_purity: 21.6%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Martin, Moore and Bennett Likely4426
    settings_parameters: "14185 x g, 34\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Olson and Sons Themselves8174
    settings_parameters: "7455 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      defense.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 31
    replicates: 2
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      design.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    temperature_celsius: 24
  - step_description: Cells were transferred with dapi stain to facilitate nothing.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 160
    replicates: 5
  - step_description: Cells were incubated with dapi stain to facilitate safe.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Thomas, Harmon and Atkinson #38783-MUST'
    concentration_or_purity: 19.4%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Jones-Kennedy #30369-PERFORM'
    concentration_or_purity: "56 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Kirk, Watkins and Hudson #35256-WORK'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Dean, Flores and Yang #23576-WAR'
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "5 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
  - equipment_name: PCR Thermocycler
    settings_parameters: "8867 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with pbs to facilitate writer.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    duration_minutes: 607
    temperature_celsius: 6
    replicates: 4
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      some.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 314
    temperature_celsius: 6
    replicates: 2
control_groups:
- control_type: Positive Control
  description: Meeting create perform most great focus either hear happy.
- control_type: Isotype Control
  description: Structure again country remember show these about often best.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
