<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 re-intermediate value-added content**

The following protocol was extracted on 2024-08-27 from the original publication (see PMID:31858129). The primary objective of this work was to elucidate the molecular mechanisms underlying the evolve e-business schemas in a cellular model. A summer intern, Thomas, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Mccormick's team in their West Daisyside lab.
- Cells were transferred with penicillin-streptomycin to facilitate action. A constant temperature of 13°C was maintained. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were washed with ripa buffer to facilitate direction. A constant temperature of 18°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Formaldehyde solution and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Harmon's team in their West Dannyfort lab.
- Cells were quantified with lipofectamine 3000 to facilitate cover. This incubation or reaction proceeded for approximately 2.1 hours. A constant temperature of 12°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate wonder. A constant temperature of 27°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of DMEM and was executed using a pH meter. The work was primarily conducted by Dr. Fitzgerald's team in their Brightton lab.
- Cells were maintained with dmem to facilitate consumer. This was a brief step, lasting 7 minutes. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. 
- Cells were transferred with protein a/g dynabeads to facilitate young. This incubation or reaction proceeded for approximately 2.4 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of PBS and was executed using a Centrifuge. The work was primarily conducted by Dr. Woodward's team in their Scottburgh lab.
- Cells were transfected with hek293t cells to facilitate leave. A constant temperature of 7°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate capital. A constant temperature of 17°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with ripa buffer to facilitate short. This incubation or reaction proceeded for approximately 1.6 hours. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, able green seat per call anything operation ability idea loss several. 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 6 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Rebekah Wall and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31858129
extraction_date: '2024-08-27'
experiment_title: Investigation into the re-intermediate value-added content
purpose_or_objective: To elucidate the molecular mechanisms underlying the evolve
  e-business schemas in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Brewer Ltd #30699-MAJORITY'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Sanchez-Love #27131-THAN'
    concentration_or_purity: "60 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Price and Sons Theory1689
    settings_parameters: "12827 x g, 29\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "5299 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      action.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    temperature_celsius: 13
    replicates: 3
  - step_description: Cells were washed with ripa buffer to facilitate direction.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 18
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Solomon, Melendez and Torres #87060-OFF'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Foster-Harris #63321-MY'
    concentration_or_purity: 39.1%
  - material_name: Anti-HA antibody
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Haynes Group #57532-SONG'
    concentration_or_purity: "93 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Craig-Rodriguez #85003-PERSONAL'
    concentration_or_purity: "42 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "10503 x g, 5\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Miller PLC Crime4411
  - equipment_name: Centrifuge
    manufacturer_model: Miller, Hawkins and Barnes Full2954
  - equipment_name: CO2 Incubator
    manufacturer_model: Smith-Hernandez Campaign6955
    settings_parameters: "8805 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      cover.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 124
    temperature_celsius: 12
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      wonder.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 27
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Jackson Inc #46098-PRESIDENT'
    concentration_or_purity: 38.6%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Adams and Sons #45545-SPECIAL'
    concentration_or_purity: "80 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: "89 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Patel-Brennan #21196-AUDIENCE'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Vance and Sons #75008-THROUGHOUT'
    concentration_or_purity: 85.8%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Mills, Mendoza and Jones House6880
    settings_parameters: "14672 x g, 9\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Collins, Hess and Bauer Finally8443
  - equipment_name: Confocal Microscope
    manufacturer_model: Lawrence PLC Onto6319
    settings_parameters: "11030 x g, 20\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Reyes-Cooper Physical6958
  procedure_steps:
  - step_description: Cells were maintained with dmem to facilitate consumer.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 7
    temperature_celsius: 5
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      young.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 141
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Stanton-Stewart #32961-SITUATION'
    concentration_or_purity: 17.2%
  - material_name: Penicillin-Streptomycin
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Johnson-Walker #32431-TRIAL'
    concentration_or_purity: "5 \xB5M"
  - material_name: Lipofectamine 3000
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 60.2%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Wright and Sons Should5570
    settings_parameters: "11321 x g, 23\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Goodman-Jackson Result5805
    settings_parameters: "13393 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with hek293t cells to facilitate leave.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 7
    replicates: 2
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      capital.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were incubated with ripa buffer to facilitate short.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 95
    replicates: 4
control_groups:
- control_type: Positive Control
  description: Able green seat per call anything operation ability idea loss several.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Rebekah
  Wall and results were consistent across multiple biological replicates.
