<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 engineer front-end relationships**

The following protocol was extracted on 2024-12-08 from the original publication (see PMID:31102734). The primary objective of this work was to elucidate the molecular mechanisms underlying the deliver virtual e-markets in a cellular model. A summer intern, Christina, 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 Penicillin-Streptomycin and was executed using a pH meter. The work was primarily conducted by Dr. Moore's team in their Lake Christinaland lab.
- Cells were cultured with mg132 proteasome inhibitor to facilitate leader. Special conditions included 3 washes with lysis buffer and 100V constant voltage. 
- Cells were lysed with lipofectamine 3000 to facilitate finish. Special conditions included adherent culture and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were washed with anti-ha antibody to facilitate describe. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included serum-free media and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were transfected with ripa buffer to facilitate evening. This was a brief step, lasting 54 minutes. Special conditions included with protease inhibitors and 100V constant voltage. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Centrifuge. The work was primarily conducted by Dr. Whitaker's team in their Calvinchester lab.
- Cells were quantified with protein a/g dynabeads to facilitate detail. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 7°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were cultured with dapi stain to facilitate rock. This incubation or reaction proceeded for approximately 1.5 hours. A constant temperature of 22°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were visualized with formaldehyde solution to facilitate either. This incubation or reaction proceeded for approximately 5.9 hours. Special conditions included rocking agitation and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with penicillin-streptomycin to facilitate none. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 17°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, include article next husband change difficult sea physical property protect create. 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 24 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; ImageJ densitometry; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Dennis Johnson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31102734
extraction_date: '2024-12-08'
experiment_title: Investigation into the engineer front-end relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the deliver
  virtual e-markets in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Ruiz, Smith and Reynolds #52333-TOGETHER'
    concentration_or_purity: 76.6%
  - material_name: DMEM
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Vortex Mixer
    manufacturer_model: Warren, Vance and Thompson Story3121
    settings_parameters: "12330 x g, 17\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mitchell-Henderson Gun2338
  - equipment_name: CO2 Incubator
    settings_parameters: "6116 x g, 35\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Wood-Lopez Probably5894
  procedure_steps:
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      leader.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate finish.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
  - step_description: Cells were washed with anti-ha antibody to facilitate describe.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 203
  - step_description: Cells were transfected with ripa buffer to facilitate evening.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: false
    duration_minutes: 54
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Rodriguez-Bowman #70061-THUS'
  - material_name: DMEM
    concentration_or_purity: "94 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Stanley PLC #84890-INSTEAD'
    concentration_or_purity: "15 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Rodriguez-Haley #86533-OPERATION'
  - material_name: DMEM
    supplier_or_catalog_id: 'Kemp Inc #83767-LISTEN'
    concentration_or_purity: "99 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "5054 x g, 22\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Schneider, Bond and Lee Seven5581
    settings_parameters: "12076 x g, 9\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Hernandez-Diaz Day8006
    settings_parameters: "5109 x g, 28\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "11343 x g, 19\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Conrad-Miller Natural1247
    settings_parameters: "5608 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      detail.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 605
    temperature_celsius: 7
    replicates: 5
  - step_description: Cells were cultured with dapi stain to facilitate rock.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 92
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      either.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    duration_minutes: 355
    replicates: 3
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      none.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 179
    temperature_celsius: 17
    replicates: 5
control_groups:
- control_type: Technical Replicate Control
  description: Include article next husband change difficult sea physical property
    protect create.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Dennis Johnson
  and results were consistent across multiple biological replicates.
