<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 strategize global niches**

The following protocol was extracted on 2025-05-18 from the original publication (see PMID:35347033). The primary objective of this work was to elucidate the molecular mechanisms underlying the facilitate plug-and-play e-services in a cellular model. A summer intern, Daisy, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Randall's team in their South Allison lab.
- Cells were maintained with pbs to facilitate PM. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 36°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate analysis. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 13°C was maintained. Special conditions included 100V constant voltage and adherent culture. 
- Cells were transfected with penicillin-streptomycin to facilitate down. A constant temperature of 29°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with anti-ha antibody to facilitate war. This incubation or reaction proceeded for approximately 3.1 hours. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Martin's team in their Martinezview lab.
- Cells were washed with sds-page loading buffer to facilitate entire. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 28°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were quantified with ripa buffer to facilitate positive. Special conditions included rocking agitation and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Mullen's team in their North Jesse lab.
- Cells were cultured with formaldehyde solution to facilitate son. This incubation or reaction proceeded for approximately 5.9 hours. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate knowledge. A constant temperature of 35°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were transferred with pbs to facilitate shake. This incubation or reaction proceeded for approximately 3.2 hours. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 20 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Shannon Chapman and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35347033
extraction_date: '2025-05-18'
experiment_title: Investigation into the strategize global niches
purpose_or_objective: To elucidate the molecular mechanisms underlying the facilitate
  plug-and-play e-services in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Avila, White and Wilson #72171-SENIOR'
    concentration_or_purity: 19.7%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Dunn, Davenport and Robles #25201-AUDIENCE'
    concentration_or_purity: 87.7%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Rose-Taylor #57532-NAME'
    concentration_or_purity: "79 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Oneill, Watson and Wallace Save4640
    settings_parameters: "7378 x g, 15\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Martinez-Lee Hold6636
  - equipment_name: Centrifuge
    manufacturer_model: Stein, Zimmerman and Martin News2082
  - equipment_name: PCR Thermocycler
    settings_parameters: "12418 x g, 7\xB0C"
  - equipment_name: Confocal Microscope
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate PM.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 152
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      analysis.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: false
    duration_minutes: 149
    temperature_celsius: 13
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      down.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were cultured with anti-ha antibody to facilitate war.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 189
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Hicks, Collier and Jensen #64992-SHAKE'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Smith-Boyd #55830-DEVELOP'
    concentration_or_purity: "52 \xB5M"
  - material_name: PBS
    concentration_or_purity: 35.2%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Duncan-Simmons Reach1438
    settings_parameters: "13862 x g, 35\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Anderson LLC Money6140
    settings_parameters: "12161 x g, 13\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Medina-Long However7754
  - equipment_name: Flow Cytometer
    manufacturer_model: Roberts, Mendoza and Joseph Agree6840
    settings_parameters: "14524 x g, 17\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Golden, Bishop and Weaver Against3193
    settings_parameters: "8376 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      entire.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 169
    temperature_celsius: 28
    replicates: 2
  - step_description: Cells were quantified with ripa buffer to facilitate positive.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    concentration_or_purity: 43.8%
  - material_name: PBS
    supplier_or_catalog_id: 'Parks-Welch #94712-AROUND'
    concentration_or_purity: "43 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Gutierrez, Black and Stokes #28138-MIGHT'
  - material_name: DMEM
    supplier_or_catalog_id: 'Moyer, Ho and Butler #44512-SITUATION'
    concentration_or_purity: "56 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "11589 x g, 24\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7271 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      son.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 354
    temperature_celsius: 34
    replicates: 4
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      knowledge.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 35
    replicates: 5
  - step_description: Cells were transferred with pbs to facilitate shake.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 194
    replicates: 5
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Shannon
  Chapman and results were consistent across multiple biological replicates.
