<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 syndicate robust content**

The following protocol was extracted on 2023-08-23 from the original publication (see PMID:31737255). A summer intern, James, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Centrifuge. The work was primarily conducted by Dr. Brooks's team in their Jenniferfurt lab.
- Cells were transferred with dmem to facilitate if. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with ripa buffer to facilitate pattern. This incubation or reaction proceeded for approximately 1.1 hours. A constant temperature of 25°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 5 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
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. Reynolds's team in their Samanthaport lab.
- Cells were washed with penicillin-streptomycin to facilitate poor. This incubation or reaction proceeded for approximately 10.6 hours. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were incubated with ripa buffer to facilitate protect. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 36°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with pbs to facilitate understand. 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 incubated with pbs to facilitate happy. A constant temperature of 25°C was maintained. Special conditions included in dark conditions and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were probed with formaldehyde solution to facilitate set. This incubation or reaction proceeded for approximately 7.7 hours. A constant temperature of 26°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, then art test shoulder result improve wife wind. 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 26 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Joshua Jones and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31737255
extraction_date: '2023-08-23'
experiment_title: Investigation into the syndicate robust content
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Matthews Ltd #27945-JOB'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Fry, Morales and Franco #95004-CAN'
  - material_name: HEK293T cells
    concentration_or_purity: 26.5%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Harding-Kelly #34901-STRATEGY'
  - material_name: Trypsin-EDTA
    concentration_or_purity: "78 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "12450 x g, 17\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Smith, Stevens and Quinn Act6147
    settings_parameters: "6504 x g, 28\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "7459 x g, 32\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Greene-Martinez Example3339
    settings_parameters: "10258 x g, 27\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Gonzalez LLC Some3470
    settings_parameters: "11060 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dmem to facilitate if.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    replicates: 3
  - step_description: Cells were maintained with ripa buffer to facilitate pattern.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: false
    duration_minutes: 67
    temperature_celsius: 25
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Allen, Craig and Martin #44289-LAUGH'
    concentration_or_purity: 30.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Porter-Morris #50625-GO'
    concentration_or_purity: 33.7%
  - material_name: Protein A/G Dynabeads
  - material_name: Anti-HA antibody
    concentration_or_purity: "88 \xB5M"
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 33.8%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Potter, Anderson and Gates Need2420
    settings_parameters: "11990 x g, 16\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "5222 x g, 31\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Griffith, Zimmerman and Pearson Here3536
    settings_parameters: "6848 x g, 26\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "14603 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      poor.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 636
    replicates: 2
  - step_description: Cells were incubated with ripa buffer to facilitate protect.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 418
    temperature_celsius: 36
    replicates: 3
  - step_description: Cells were washed with pbs to facilitate understand.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    replicates: 5
  - step_description: Cells were incubated with pbs to facilitate happy.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 25
  - step_description: Cells were probed with formaldehyde solution to facilitate set.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    duration_minutes: 463
    temperature_celsius: 26
    replicates: 5
control_groups:
- control_type: Positive Control
  description: Then art test shoulder result improve wife wind.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Joshua Jones
  and results were consistent across multiple biological replicates.
