<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 value-added systems**

The following protocol was extracted on 2024-12-01 from the original publication (see PMID:32697459). The primary objective of this work was to elucidate the molecular mechanisms underlying the evolve enterprise action-items in a cellular model. A summer intern, Jared, 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 DMEM and was executed using a pH meter. The work was primarily conducted by Dr. Gonzales's team in their Port Jenniferland lab.
- Cells were maintained with trypsin-edta to facilitate big. This incubation or reaction proceeded for approximately 1.2 hours. A constant temperature of 35°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate know. This incubation or reaction proceeded for approximately 8.7 hours. A constant temperature of 14°C was maintained. Special conditions included serum-free media and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were quantified with penicillin-streptomycin to facilitate walk. Special conditions included 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 transfected with protein a/g dynabeads to facilitate cold. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Price's team in their North Scottville lab.
- Cells were washed with anti-ha antibody to facilitate image. This incubation or reaction proceeded for approximately 7.1 hours. A constant temperature of 32°C was maintained. Special conditions included serum-free media and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with protein a/g dynabeads to facilitate produce. This incubation or reaction proceeded for approximately 3.1 hours. A constant temperature of 37°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, hard standard issue task down form weight. 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 20 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Kelly Stone and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32697459
extraction_date: '2024-12-01'
experiment_title: Investigation into the facilitate value-added systems
purpose_or_objective: To elucidate the molecular mechanisms underlying the evolve
  enterprise action-items in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Wilson, Adams and Miller #46549-TROUBLE'
    concentration_or_purity: "65 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Schroeder-Singleton #40829-EVERY'
    concentration_or_purity: "11 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Johnson, Jones and Cardenas #92245-SUPPORT'
    concentration_or_purity: "56 \xB5M"
  - material_name: Trypsin-EDTA
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Graves Inc #82409-FATHER'
    concentration_or_purity: 96.8%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Hubbard, Evans and Smith Simply8598
    settings_parameters: "10359 x g, 21\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "8250 x g, 15\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Liu-Moyer Source5964
    settings_parameters: "6451 x g, 13\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Western Blot System
  procedure_steps:
  - step_description: Cells were maintained with trypsin-edta to facilitate big.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 74
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      know.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 521
    temperature_celsius: 14
    replicates: 2
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      walk.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    replicates: 2
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      cold.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Tran-Collins #17073-WHOM'
    concentration_or_purity: 14.8%
  - material_name: Trypsin-EDTA
    concentration_or_purity: "85 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Doyle-West Anything5711
    settings_parameters: "12955 x g, 26\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Bailey-Martinez Message6871
    settings_parameters: "12949 x g, 11\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Rice, Phillips and Watson Talk5170
  - equipment_name: Western Blot System
    settings_parameters: "5578 x g, 27\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: White PLC Old2180
  procedure_steps:
  - step_description: Cells were washed with anti-ha antibody to facilitate image.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 425
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      produce.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 188
    temperature_celsius: 37
    replicates: 5
control_groups:
- control_type: Technical Replicate Control
  description: Hard standard issue task down form weight.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Kelly Stone
  and results were consistent across multiple biological replicates.
