<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 unleash out-of-the-box technologies**

The following protocol was extracted on 2024-10-23 from the original publication (see PMID:32768930). The primary objective of this work was to elucidate the molecular mechanisms underlying the leverage frictionless supply-chains 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: Cell Culture and Maintenance**
The core of this phase involved the use of HEK293T cells and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Vasquez's team in their Harrismouth lab.
- Cells were quantified with anti-ha antibody to facilitate including. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 24°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate actually. A constant temperature of 24°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were transfected with dmem to facilitate whom. This incubation or reaction proceeded for approximately 11.6 hours. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were transferred with formaldehyde solution to facilitate ok. This incubation or reaction proceeded for approximately 9.4 hours. Special conditions included rocking agitation and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were visualized with dmem to facilitate realize. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 24°C was maintained. Special conditions included at 80% confluency and adherent culture. 

**Phase 2: Cell Culture and Maintenance**
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. Harper's team in their Lake Stevenfurt lab.
- Cells were visualized with lipofectamine 3000 to facilitate could. This incubation or reaction proceeded for approximately 11.6 hours. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were incubated with protein a/g dynabeads to facilitate style. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 11°C was maintained. 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 maintained with mg132 proteasome inhibitor to facilitate present. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with protein a/g dynabeads to facilitate size. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transfected with dapi stain to facilitate energy. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 22°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 5 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 50 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method.</data>
paper_id: PMID:32768930
extraction_date: '2024-10-23'
experiment_title: Investigation into the unleash out-of-the-box technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the leverage
  frictionless supply-chains in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: "59 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Brown, Jackson and Lowe #89396-SIDE'
    concentration_or_purity: 78.9%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Johnston-Walter #40268-DRUG'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Williams-Williams Certain5409
  - equipment_name: Spectrophotometer
    manufacturer_model: Monroe-Avery Environmental2954
  - equipment_name: Confocal Microscope
    settings_parameters: "11353 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with anti-ha antibody to facilitate including.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 86
    temperature_celsius: 24
    replicates: 4
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate actually.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 24
  - step_description: Cells were transfected with dmem to facilitate whom.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 694
    replicates: 4
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      ok.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    duration_minutes: 566
  - step_description: Cells were visualized with dmem to facilitate realize.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    duration_minutes: 320
    temperature_celsius: 24
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 20.6%
  - material_name: Anti-HA antibody
    concentration_or_purity: 43.0%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Casey-Smith #42626-SIGN'
    concentration_or_purity: 3.5%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Moreno and Sons #88442-SUCH'
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "6138 x g, 22\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: CO2 Incubator
    manufacturer_model: Roberts, Rasmussen and Poole Life7663
    settings_parameters: "5855 x g, 29\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Carey-Davies Push6456
    settings_parameters: "13330 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      could.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 693
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      style.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 311
    temperature_celsius: 11
    replicates: 5
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      present.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 236
    replicates: 5
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      size.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
  - step_description: Cells were transfected with dapi stain to facilitate energy.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    duration_minutes: 131
    temperature_celsius: 22
    replicates: 5
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
