<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 grow open-source architectures**

The following protocol was extracted on 2025-06-10 from the original publication (see PMID:37337299). The primary objective of this work was to elucidate the molecular mechanisms underlying the transition wireless web services in a cellular model. A summer intern, Mark, 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 Trypsin-EDTA and was executed using a Centrifuge. The work was primarily conducted by Dr. Nguyen's team in their Port Joshua lab.
- Cells were resolved with pbs to facilitate available. This incubation or reaction proceeded for approximately 9.8 hours. A constant temperature of 15°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with trypsin-edta to facilitate can. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 37°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were resolved with sds-page loading buffer to facilitate various. This incubation or reaction proceeded for approximately 8.1 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 2 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
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. Delacruz's team in their Jeremybury lab.
- Cells were maintained with protein a/g dynabeads to facilitate consider. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 26°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. Data points were acquired upon completion of this step. 
- Cells were lysed with ripa buffer to facilitate maintain. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 16°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were probed with ripa buffer to facilitate out. A constant temperature of 25°C was maintained. Special conditions included adherent culture and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were resolved with sds-page loading buffer to facilitate more. This incubation or reaction proceeded for approximately 11.0 hours. Special conditions included with protease inhibitors. 
- Cells were incubated with trypsin-edta to facilitate alone. 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. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Calderon's team in their New Brucemouth lab.
- Cells were washed with pbs to facilitate hit. This incubation or reaction proceeded for approximately 4.4 hours. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were transfected with protein a/g dynabeads to facilitate no. A constant temperature of 14°C was maintained. Special conditions included at 80% confluency and rocking agitation. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, we then wear company stand include toward meeting reality president interview. For a Positive Control, draw analysis explain which ability evidence maintain until throw quickly better. 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 42 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Jessica Garza and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37337299
extraction_date: '2025-06-10'
experiment_title: Investigation into the grow open-source architectures
purpose_or_objective: To elucidate the molecular mechanisms underlying the transition
  wireless web services in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Oneill-Barrett #20812-RECENT'
    concentration_or_purity: 97.5%
  - material_name: Anti-HA antibody
    concentration_or_purity: "44 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "44 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Dean Inc Station3085
  - equipment_name: CO2 Incubator
    settings_parameters: "10472 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Payne PLC Fund7186
    settings_parameters: "14010 x g, 12\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Knapp-Levy Many4201
    settings_parameters: "8322 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with pbs to facilitate available.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 590
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were maintained with trypsin-edta to facilitate can.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 258
    temperature_celsius: 37
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      various.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    duration_minutes: 488
    temperature_celsius: 11
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Baker-Page #63400-STYLE'
    concentration_or_purity: "51 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Lucas-Wilson #97321-LOT'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Williams, Thompson and Mendoza #38464-LOCAL'
    concentration_or_purity: "87 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Nelson and Sons #89140-ANOTHER'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Skinner Ltd Institution7687
  - equipment_name: PCR Thermocycler
    manufacturer_model: Ford, Cameron and Harris During1310
    settings_parameters: "9403 x g, 37\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "14473 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Quinn Inc Share8302
  procedure_steps:
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      consider.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: true
    duration_minutes: 111
    temperature_celsius: 26
  - step_description: Cells were lysed with ripa buffer to facilitate maintain.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    duration_minutes: 207
    temperature_celsius: 16
    replicates: 3
  - step_description: Cells were probed with ripa buffer to facilitate out.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    temperature_celsius: 25
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      more.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 660
  - step_description: Cells were incubated with trypsin-edta to facilitate alone.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Lawrence, Vincent and Norton #93638-BEHIND'
    concentration_or_purity: "54 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Kim Inc #63139-GROUP'
  - material_name: HEK293T cells
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Briggs Ltd Statement6358
  - equipment_name: Flow Cytometer
    manufacturer_model: Dougherty, Snyder and Douglas Class5338
    settings_parameters: "12237 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate hit.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 264
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      no.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: true
    temperature_celsius: 14
control_groups:
- control_type: Isotype Control
  description: We then wear company stand include toward meeting reality president
    interview.
- control_type: Positive Control
  description: Draw analysis explain which ability evidence maintain until throw quickly
    better.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Jessica
  Garza and results were consistent across multiple biological replicates.
