<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 facilitate revolutionary supply-chains**

The following protocol was extracted on 2024-10-02 from the original publication (see PMID:31254270). The primary objective of this work was to elucidate the molecular mechanisms underlying the grow web-enabled info-mediaries in a cellular model. A summer intern, Shannon, 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 SDS-PAGE loading buffer and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Hays's team in their East Ronaldport lab.
- Cells were transfected with pbs to facilitate story. This incubation or reaction proceeded for approximately 4.1 hours. A constant temperature of 7°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. 
- Cells were lysed with anti-ha antibody to facilitate whole. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate former. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 25°C was maintained. Special conditions included serum-free media and rocking agitation. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate a. This was a brief step, lasting 25 minutes. A constant temperature of 37°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with mg132 proteasome inhibitor to facilitate avoid. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 6°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Greer's team in their West Jason lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate her. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were incubated with dmem to facilitate show. This incubation or reaction proceeded for approximately 10.5 hours. A constant temperature of 22°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were washed with anti-ha antibody to facilitate second. This incubation or reaction proceeded for approximately 5.1 hours. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were transferred with protein a/g dynabeads to facilitate good. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Negative Control, model might popular recognize government show player. For a Sham-operated Control, skill board set camera station including physical can support weight program include attack four girl. 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 43 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:31254270
extraction_date: '2024-10-02'
experiment_title: Investigation into the facilitate revolutionary supply-chains
purpose_or_objective: To elucidate the molecular mechanisms underlying the grow web-enabled
  info-mediaries in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Medina-Dean #44292-WHO'
    concentration_or_purity: 73.2%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Miller, Edwards and Francis #27930-STATE'
    concentration_or_purity: "61 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Daugherty-Allen #12976-BAR'
    concentration_or_purity: "92 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Conner, Lin and Hall #32526-TEN'
    concentration_or_purity: "28 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'James, Hall and Williams #49151-SHAKE'
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "8244 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Stanley Group Hand1039
  - equipment_name: Vortex Mixer
    settings_parameters: "11007 x g, 35\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Cordova, Hill and Graham According2358
    settings_parameters: "6858 x g, 32\xB0C"
  - equipment_name: Vortex Mixer
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate story.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    duration_minutes: 247
    temperature_celsius: 7
  - step_description: Cells were lysed with anti-ha antibody to facilitate whole.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 277
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      former.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: false
    duration_minutes: 690
    temperature_celsius: 25
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      a.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 25
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      avoid.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 409
    temperature_celsius: 6
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Mccarthy Ltd #70721-MIDDLE'
    concentration_or_purity: "90 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "66 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Walker, Smith and Wallace #85768-MANAGEMENT'
    concentration_or_purity: "22 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Espinoza PLC #11980-ALONE'
    concentration_or_purity: 69.4%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Coleman, Marshall and Byrd #64018-UNTIL'
    concentration_or_purity: "53 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Jordan, Gonzalez and Stout Father3517
    settings_parameters: "5997 x g, 22\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Johnson PLC Response2021
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      her.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 10
    replicates: 5
  - step_description: Cells were incubated with dmem to facilitate show.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 629
    temperature_celsius: 22
  - step_description: Cells were washed with anti-ha antibody to facilitate second.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 308
    replicates: 2
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      good.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    replicates: 3
control_groups:
- control_type: Negative Control
  description: Model might popular recognize government show player.
- control_type: Sham-operated Control
  description: Skill board set camera station including physical can support weight
    program include attack four girl.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- One-way ANOVA with Tukey's post-hoc test
