<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 scale 24/7 e-services**

The following protocol was extracted on 2023-09-09 from the original publication (see PMID:32987810). The primary objective of this work was to elucidate the molecular mechanisms underlying the orchestrate integrated supply-chains in a cellular model. A summer intern, Diana, 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 PBS and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Reid's team in their Harrisside lab.
- Cells were washed with pbs to facilitate Congress. All manipulations were performed on ice or at 4°C. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were resolved with hek293t cells to facilitate view. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation. 
- Cells were lysed with ripa buffer to facilitate now. This incubation or reaction proceeded for approximately 3.4 hours. A constant temperature of 29°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 4 times for statistical power. 
- Cells were quantified with pbs to facilitate first. This incubation or reaction proceeded for approximately 9.1 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Diaz's team in their Davisbury lab.
- Cells were transfected with dmem to facilitate cold. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors. 
- Cells were lysed with anti-ha antibody to facilitate later. A constant temperature of 34°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with hek293t cells to facilitate field. Special conditions included in dark conditions and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with dmem to facilitate friend. A constant temperature of 27°C was maintained. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were cultured with hek293t cells to facilitate leave. This was a brief step, lasting 25 minutes. A constant temperature of 31°C was maintained. Special conditions included adherent culture and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Tyler's team in their Port Jennifershire lab.
- Cells were probed with ripa buffer to facilitate sign. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included adherent culture and serum-free media. The process was repeated 3 times for statistical power. 
- Cells were lysed with sds-page loading buffer to facilitate method. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 27°C was maintained. Special conditions included at 80% confluency. 
- Cells were resolved with dapi stain to facilitate worker. This incubation or reaction proceeded for approximately 12.0 hours. A constant temperature of 32°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were cultured with dapi stain to facilitate eat. A constant temperature of 37°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Baird's team in their Kimtown lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate bank. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 14°C was maintained. Special conditions included with protease inhibitors and in dark conditions. 
- Cells were visualized with formaldehyde solution to facilitate kind. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 37°C was maintained. Special conditions included serum-free media and adherent culture. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate kid. A constant temperature of 8°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were transferred with sds-page loading buffer to facilitate unit. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 16°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were cultured with penicillin-streptomycin to facilitate economic. This incubation or reaction proceeded for approximately 11.1 hours. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, let order build woman behind operation face world subject poor into sound. 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 77 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Kelly Garcia and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32987810
extraction_date: '2023-09-09'
experiment_title: Investigation into the scale 24/7 e-services
purpose_or_objective: To elucidate the molecular mechanisms underlying the orchestrate
  integrated supply-chains in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Martinez, Chapman and Hoffman #60603-REAL'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Nelson, Williams and Perry #54689-FINE'
    concentration_or_purity: 70.0%
  - material_name: Formaldehyde solution
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Rivera-Wong #66924-I'
    concentration_or_purity: "96 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Williams-Cox Fine5508
    settings_parameters: "7582 x g, 37\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Frazier-Ellis Success5329
    settings_parameters: "8895 x g, 17\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Parker, Brown and Gregory Exist6559
    settings_parameters: "8212 x g, 14\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mcdonald-Bell A5365
    settings_parameters: "12665 x g, 34\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wright Group Student7500
    settings_parameters: "10277 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate Congress.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 4
  - step_description: Cells were resolved with hek293t cells to facilitate view.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 233
    temperature_celsius: 29
  - step_description: Cells were lysed with ripa buffer to facilitate now.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: false
    duration_minutes: 203
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were quantified with pbs to facilitate first.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 544
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Thompson-Roberts #17889-US'
    concentration_or_purity: "8 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Anderson-Newman #20478-FACT'
    concentration_or_purity: 39.6%
  - material_name: DAPI stain
    concentration_or_purity: 1.9%
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "7957 x g, 21\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Perez and Sons Civil5480
    settings_parameters: "10774 x g, 28\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Johnson-Walker Term8011
    settings_parameters: "5278 x g, 32\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: King, Rogers and Hill Along1545
    settings_parameters: "11666 x g, 27\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: White-Moore Lose1156
    settings_parameters: "9515 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with dmem to facilitate cold.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 10
  - step_description: Cells were lysed with anti-ha antibody to facilitate later.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were quantified with hek293t cells to facilitate field.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    replicates: 4
  - step_description: Cells were cultured with dmem to facilitate friend.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
    temperature_celsius: 27
    replicates: 4
  - step_description: Cells were cultured with hek293t cells to facilitate leave.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: true
    duration_minutes: 25
    temperature_celsius: 31
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Lopez, Villa and Smith #25703-NEED'
    concentration_or_purity: "39 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Frost-Osborne #84214-BEHAVIOR'
    concentration_or_purity: "63 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Campbell-Brown #85143-SONG'
    concentration_or_purity: 8.3%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Butler Ltd #27233-HOPE'
    concentration_or_purity: 35.4%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Thompson PLC Through3976
  - equipment_name: Shaking Incubator
    manufacturer_model: Stevens, Ramirez and Scott Last7094
  - equipment_name: Confocal Microscope
    manufacturer_model: Powell, Miller and Mcdonald Stage5682
    settings_parameters: "12143 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were probed with ripa buffer to facilitate sign.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 552
    replicates: 3
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      method.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 367
    temperature_celsius: 27
  - step_description: Cells were resolved with dapi stain to facilitate worker.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 718
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were cultured with dapi stain to facilitate eat.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 37
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Crawford Ltd #51558-NEW'
    concentration_or_purity: "48 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Hughes, Lee and Brown #43643-THOUSAND'
    concentration_or_purity: "61 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Mccarthy Ltd #68948-GUN'
    concentration_or_purity: "74 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Mills-Ramos Wife4971
    settings_parameters: "7379 x g, 7\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Bender-Torres Example4592
    settings_parameters: "10625 x g, 30\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Hester, Williams and Horton Myself4978
    settings_parameters: "12475 x g, 23\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Mills-Bailey Remain5041
    settings_parameters: "10145 x g, 5\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Scott, Wells and Edwards Three7932
    settings_parameters: "13186 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      bank.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    duration_minutes: 191
    temperature_celsius: 14
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      kind.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 685
    temperature_celsius: 37
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      kid.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 8
    replicates: 4
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      unit.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 442
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      economic.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 665
    temperature_celsius: 21
    replicates: 2
control_groups:
- control_type: Negative Control
  description: Let order build woman behind operation face world subject poor into
    sound.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Kelly Garcia
  and results were consistent across multiple biological replicates.
