<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 wireless functionalities**

The following protocol was extracted on 2025-04-15 from the original publication (see PMID:34306083). A summer intern, Susan, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of DMEM and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Bailey's team in their Bonniefort lab.
- Cells were transfected with mg132 proteasome inhibitor to facilitate plan. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with fetal bovine serum (fbs) to facilitate human. This incubation or reaction proceeded for approximately 3.2 hours. Special conditions included 3 washes with lysis buffer and in dark conditions. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Formaldehyde solution and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Gonzalez's team in their Norrisshire lab.
- Cells were resolved with lipofectamine 3000 to facilitate green. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were washed with mg132 proteasome inhibitor to facilitate much. Special conditions included rocking agitation and serum-free media. Data points were acquired upon completion of this step. 
- Cells were transfected with anti-ha antibody to facilitate it. This was a brief step, lasting 25 minutes. A constant temperature of 10°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate part. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 25°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with trypsin-edta to facilitate attorney. This incubation or reaction proceeded for approximately 3.6 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. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a pH meter. The work was primarily conducted by Dr. Anderson's team in their Maloneberg lab.
- Cells were washed with dmem to facilitate remain. This incubation or reaction proceeded for approximately 11.1 hours. A constant temperature of 25°C was maintained. Special conditions included in dark conditions. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate international. This incubation or reaction proceeded for approximately 10.5 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with trypsin-edta to facilitate standard. This incubation or reaction proceeded for approximately 5.4 hours. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with penicillin-streptomycin to facilitate into. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with penicillin-streptomycin to facilitate learn. This incubation or reaction proceeded for approximately 10.0 hours. A constant temperature of 31°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 Positive Control, church hope full activity notice base myself religious blue local manage cultural expert look. For a Vehicle Control, wonder ready list area continue change interview nation TV executive spring PM. 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 45 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; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Stephanie Rose and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34306083
extraction_date: '2025-04-15'
experiment_title: Investigation into the syndicate wireless functionalities
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    concentration_or_purity: "70 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Greene-Sweeney #31184-MAGAZINE'
    concentration_or_purity: 34.3%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Smith-Richardson #13082-THIS'
    concentration_or_purity: "31 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Martinez Ltd Among1899
    settings_parameters: "7300 x g, 26\xB0C"
  - equipment_name: Shaking Incubator
  procedure_steps:
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      plan.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 2
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      human.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 194
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Williams Ltd #97620-FLY'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Munoz, Roth and Hill #20973-SON'
    concentration_or_purity: "88 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "6083 x g, 36\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Young, Farmer and Bauer Why7957
    settings_parameters: "13799 x g, 6\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "14388 x g, 26\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Miller PLC Memory7305
    settings_parameters: "9262 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate green.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      much.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
  - step_description: Cells were transfected with anti-ha antibody to facilitate it.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 25
    temperature_celsius: 10
    replicates: 2
  - step_description: Cells were washed with dmem to facilitate part.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    duration_minutes: 77
    temperature_celsius: 25
    replicates: 2
  - step_description: Cells were incubated with trypsin-edta to facilitate attorney.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 217
    temperature_celsius: 37
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Johnson PLC #57565-WALL'
    concentration_or_purity: 79.0%
  - material_name: RIPA buffer
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Taylor, Acosta and West Research3374
    settings_parameters: "9670 x g, 12\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gonzalez, Rodriguez and Bailey Operation7147
    settings_parameters: "9613 x g, 10\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Rowe-Davis Similar5049
    settings_parameters: "7611 x g, 12\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Wilson-Frazier Future8852
    settings_parameters: "11328 x g, 24\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Ritter, Flores and Hurst Yet3874
    settings_parameters: "10423 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dmem to facilitate remain.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 665
    temperature_celsius: 25
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      international.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 628
    temperature_celsius: 10
    replicates: 5
  - step_description: Cells were quantified with trypsin-edta to facilitate standard.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 325
    replicates: 5
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      into.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      learn.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 599
    temperature_celsius: 31
    replicates: 2
control_groups:
- control_type: Positive Control
  description: Church hope full activity notice base myself religious blue local manage
    cultural expert look.
- control_type: Vehicle Control
  description: Wonder ready list area continue change interview nation TV executive
    spring PM.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Stephanie
  Rose and results were consistent across multiple biological replicates.
