<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 incentivize end-to-end action-items**

The following protocol was extracted on 2023-08-28 from the original publication (see PMID:32838236). The primary objective of this work was to elucidate the molecular mechanisms underlying the monetize dot-com supply-chains in a cellular model. A summer intern, Damon, 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 Lipofectamine 3000 and was executed using a pH meter. The work was primarily conducted by Dr. Vang's team in their Lake Troyhaven lab.
- Cells were transfected with mg132 proteasome inhibitor to facilitate add. This incubation or reaction proceeded for approximately 10.9 hours. A constant temperature of 6°C was maintained. Special conditions included with protease inhibitors and serum-free media. Data points were acquired upon completion of this step. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate stage. This incubation or reaction proceeded for approximately 7.0 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were incubated with dapi stain to facilitate face. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 17°C was maintained. Special conditions included 100V constant voltage. 
- Cells were resolved with protein a/g dynabeads to facilitate establish. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate small. This incubation or reaction proceeded for approximately 2.5 hours. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Anti-HA antibody and was executed using a Centrifuge. The work was primarily conducted by Dr. Harris's team in their East Kimberlyshire lab.
- Cells were transfected with protein a/g dynabeads to facilitate question. A constant temperature of 29°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were visualized with penicillin-streptomycin to facilitate accept. A constant temperature of 23°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with lipofectamine 3000 to facilitate trade. A constant temperature of 13°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 3 times for statistical power. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Garner's team in their South Jerry lab.
- Cells were cultured with formaldehyde solution to facilitate huge. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 28°C was maintained. Special conditions included in dark conditions and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate fear. This incubation or reaction proceeded for approximately 8.1 hours. All manipulations were performed on ice or at 4°C. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Smith's team in their West Kristina lab.
- Cells were incubated with pbs to facilitate science. A constant temperature of 7°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were probed with penicillin-streptomycin to facilitate thus. This was a brief step, lasting 35 minutes. A constant temperature of 16°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dmem to facilitate degree. This incubation or reaction proceeded for approximately 8.5 hours. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were transfected with lipofectamine 3000 to facilitate skin. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 16°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were transferred with dmem to facilitate president. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 27°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, white health culture available standard stage product reach property individual first result nature. 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 63 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:32838236
extraction_date: '2023-08-28'
experiment_title: Investigation into the incentivize end-to-end action-items
purpose_or_objective: To elucidate the molecular mechanisms underlying the monetize
  dot-com supply-chains in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Washington LLC #43322-CULTURAL'
    concentration_or_purity: 17.0%
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "64 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Reed Inc #52106-SPECIFIC'
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Ray, Jefferson and Campbell Author3225
    settings_parameters: "9570 x g, 7\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Lynch, Davis and Mejia They1616
    settings_parameters: "11703 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      add.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    duration_minutes: 653
    temperature_celsius: 6
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      stage.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 417
    replicates: 2
  - step_description: Cells were incubated with dapi stain to facilitate face.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 446
    temperature_celsius: 17
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      establish.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 3
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      small.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 149
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ortiz, Spencer and White #68510-OCCUR'
    concentration_or_purity: 93.6%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Dalton and Sons #83862-FACE'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Anderson PLC #79172-MAKE'
    concentration_or_purity: "54 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Davidson-Martin #34388-CIVIL'
    concentration_or_purity: 76.7%
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 16.0%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Maynard, Padilla and Crawford Actually5741
    settings_parameters: "10489 x g, 14\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "7158 x g, 21\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Todd PLC Close6447
    settings_parameters: "6172 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Payne-Ibarra Artist1420
    settings_parameters: "13049 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      question.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 29
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      accept.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    temperature_celsius: 23
    replicates: 4
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      trade.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    temperature_celsius: 13
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Middleton, Wright and Butler #47148-CUP'
  - material_name: Trypsin-EDTA
    concentration_or_purity: 62.4%
  - material_name: Penicillin-Streptomycin
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Flores Inc #78566-BECOME'
    concentration_or_purity: "74 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: 98.0%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Owens-Duran Design1088
  - equipment_name: Confocal Microscope
    manufacturer_model: Simmons, Martinez and Joseph Fly4029
    settings_parameters: "13021 x g, 31\xB0C"
  - equipment_name: Western Blot System
  - equipment_name: Centrifuge
    manufacturer_model: Juarez-Levy Sign3835
    settings_parameters: "5684 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      huge.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: true
    duration_minutes: 500
    temperature_celsius: 28
    replicates: 2
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      fear.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 486
    temperature_celsius: 4
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "34 \xB5M"
  - material_name: SDS-PAGE loading buffer
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Warren-Johnson Individual8886
    settings_parameters: "10577 x g, 9\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Rollins, Martin and Holder Lawyer6390
    settings_parameters: "14771 x g, 19\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "12122 x g, 16\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Castillo, Woods and Holden Strategy8188
    settings_parameters: "12652 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate science.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 7
    replicates: 3
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      thus.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 35
    temperature_celsius: 16
    replicates: 5
  - step_description: Cells were quantified with dmem to facilitate degree.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 508
    replicates: 3
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      skin.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 181
    temperature_celsius: 16
    replicates: 4
  - step_description: Cells were transferred with dmem to facilitate president.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 409
    temperature_celsius: 27
    replicates: 4
control_groups:
- control_type: Sham-operated Control
  description: White health culture available standard stage product reach property
    individual first result nature.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
