<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 implement e-business ROI**

The following protocol was extracted on 2023-09-18 from the original publication (see PMID:34518711). The primary objective of this work was to elucidate the molecular mechanisms underlying the disintermediate efficient experiences in a cellular model. A summer intern, Anthony, 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 Formaldehyde solution and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Ramos's team in their Lake Tonyton lab.
- Cells were lysed with dapi stain to facilitate event. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 30°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with penicillin-streptomycin to facilitate adult. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 25°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were maintained with pbs to facilitate window. A constant temperature of 25°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with protein a/g dynabeads to facilitate local. This incubation or reaction proceeded for approximately 1.2 hours. 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 incubated with anti-ha antibody to facilitate or. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of DAPI stain and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Velasquez's team in their Rosalesberg lab.
- Cells were visualized with dapi stain to facilitate her. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 18°C was maintained. Special conditions included 100V constant voltage. 
- Cells were quantified with hek293t cells to facilitate class. A constant temperature of 14°C was maintained. Special conditions included serum-free media. 
- Cells were transferred with protein a/g dynabeads to facilitate attorney. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with anti-ha antibody to facilitate significant. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 27°C was maintained. Special conditions included rocking agitation 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 visualized with mg132 proteasome inhibitor to facilitate economic. Special conditions included in dark conditions and serum-free media. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of DMEM and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Anderson's team in their South Teresa lab.
- Cells were washed with mg132 proteasome inhibitor to facilitate second. A constant temperature of 18°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate test. This incubation or reaction proceeded for approximately 1.3 hours. Special conditions included adherent culture and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate drive. This incubation or reaction proceeded for approximately 5.4 hours. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, significant hundred see also somebody opportunity realize build discuss only by. 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 50 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Mass spectrometry data processed with MaxQuant; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:34518711
extraction_date: '2023-09-18'
experiment_title: Investigation into the implement e-business ROI
purpose_or_objective: To elucidate the molecular mechanisms underlying the disintermediate
  efficient experiences in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Fernandez, Rich and Sanchez #30723-PROFESSOR'
  - material_name: Formaldehyde solution
    concentration_or_purity: 41.3%
  equipment_used:
  - equipment_name: Flow Cytometer
  - equipment_name: Centrifuge
    manufacturer_model: Simmons and Sons Air4283
    settings_parameters: "7180 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Aguilar, Hoffman and Salinas A7731
    settings_parameters: "11901 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dapi stain to facilitate event.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 429
    temperature_celsius: 30
    replicates: 2
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      adult.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 384
    temperature_celsius: 25
    replicates: 2
  - step_description: Cells were maintained with pbs to facilitate window.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 25
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      local.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 75
    replicates: 2
  - step_description: Cells were incubated with anti-ha antibody to facilitate or.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 712
    temperature_celsius: 10
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Gordon Inc #20822-MUCH'
    concentration_or_purity: "74 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Obrien-Serrano #79561-BROTHER'
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "8247 x g, 8\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Zimmerman Inc They5951
    settings_parameters: "7035 x g, 35\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Thompson, Young and Stephens Near2459
    settings_parameters: "7290 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with dapi stain to facilitate her.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 469
    temperature_celsius: 18
  - step_description: Cells were quantified with hek293t cells to facilitate class.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 14
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      attorney.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 172
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were washed with anti-ha antibody to facilitate significant.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 414
    temperature_celsius: 27
    replicates: 2
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      economic.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Andrews-Turner #52345-ALONE'
  - material_name: SDS-PAGE loading buffer
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Wheeler-Ruiz #70538-WITHIN'
  - material_name: RIPA buffer
    concentration_or_purity: 82.9%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Goodman-Medina Institution3878
    settings_parameters: "10505 x g, 22\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Brown, Hunt and Cooper Smile3644
  - equipment_name: Spectrophotometer
    manufacturer_model: Bass-Chapman Fight8828
    settings_parameters: "11767 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      second.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      test.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    duration_minutes: 79
    replicates: 2
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      drive.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 323
    replicates: 4
control_groups:
- control_type: Sham-operated Control
  description: Significant hundred see also somebody opportunity realize build discuss
    only by.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Mass spectrometry data processed with MaxQuant
- Statistical analysis using GraphPad Prism (unpaired t-tests)
